KR102274232B1 - A composition comprising neuro-targeted proteins - Google Patents
A composition comprising neuro-targeted proteins Download PDFInfo
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Abstract
본 발명은 BoNTs의 RBD 또는 A2 시퀀스와 EEPs를 포함함으로써 뉴런을 특이적 인식하고 소포 탈출이 가능한 조성물, 조성물의 제조방법 및 이를 이용한 약학적 조성물, 뉴런 표시용 조성물, 및 화장료 조성물 등에 관한 것이다. 본 발명에서 제조된 재조합 조성물은 생체뉴런 내·외부에서 특이적으로 생리활성 또는 표시 기능을 갖으면서도 단백질의 구조를 단순화하여 분자량을 줄인 것으로 생산과정의 단순화를 도모할 수 있어 경제적이고, 작은 분자량에 의해 항체, 항원 반응으로부터 더 자유로울 수 있으며, 나아가, 화장료로 사용되는 경우 분자의 크기가 작아 흡수율이 증대된다는 이점이 있어 관련된 의료사업분야 및 화장품 산업 분야 등에 다양하게 이용될 수 있을 것으로 기대된다.The present invention relates to a composition capable of specifically recognizing neurons and vesicle escape by including the RBD or A2 sequence of BoNTs and EEPs, a method for preparing the composition, and a pharmaceutical composition using the same, a composition for displaying neurons, a cosmetic composition, and the like. The recombinant composition prepared in the present invention has a specific physiological activity or display function inside and outside biological neurons, while simplifying the structure of the protein and reducing the molecular weight, thereby simplifying the production process, which is economical and has a small molecular weight. It can be more free from antibody and antigen reactions by means of an antibody and antigen reaction, and further, when used as a cosmetic, it has the advantage that the absorption rate is increased due to the small size of the molecule, so it is expected to be variously used in the related medical business field and the cosmetic industry field.
Description
본 발명은 신경세포 표적용 단백질 조성물에 관한 것으로, 보다 자세하게는, 신경세포(뉴런)을 특이적으로 인식하는 재조합 단백질 조성물에 관한 것으로, 신경-활성 물질에 뉴런을 특이적으로 인식하고 뉴런 내 소포를 탈출하는 서열을 결합시킴으로써, 신경-활성 물질 재조합 단백질이 뉴런을 특이적으로 인식하고, 뉴런 내의 세포질로 이동하여 활성을 나타내는 방법, 이를 포함하는 조성물, 제조 방법, 및 이의 용도 등에 관한 것이다.The present invention relates to a protein composition for targeting a nerve cell, and more particularly, to a recombinant protein composition for specifically recognizing a nerve cell (neuron), which specifically recognizes a neuron by a neuron-active substance and vesicles within the neuron It relates to a method in which a neuron-active substance recombinant protein specifically recognizes a neuron by binding a sequence that escapes the neuron, and moves to the cytoplasm in the neuron to exhibit activity, a composition comprising the same, a manufacturing method, and uses thereof, and the like.
보툴리눔 신경독소(botulinum neurotoxin, BoNT)는 신경세포(뉴런, neuron)를 인식하여 결합하는 기능을 가진 부분(receptor binding domain, RBD)을 포함하고 있다. 상기 BoNT 단백질의 RBD는 시냅스 전막의 당지질과 결합하는 아미노산 시퀀스와 시냅스 전막 상에 존재하는 단백질과 특이적으로 결합하는 아미노산 시퀀스 중 전자만을 포함하거나 모두 포함하고 있다. 한편, A타입 BoNT(BoNT/A)의 엔도사이토시스(endocytosis)는 RBD 내의 A2 펩티드(1040-1053, 13개의 특정 아미노산으로 구성)가 시냅스 전막 상에 존재하는 시냅스 소포 당단백질 2C(synaptic vesicle glycoprotein 2C, SV2C)의 루미날 도메인(luminal domain)에 특이적으로 결합함으로써 가능하다 알려져 있으며, 상기 결합은 다른(B-G) 타입의 BoNT와 시냅스 전막 단백질과의 결합에 비해 강력할 것으로 예상되고 있다.Botulinum neurotoxin (BoNT) includes a receptor binding domain (RBD) that recognizes and binds nerve cells (neurons). The RBD of the BoNT protein contains only the former or all of the amino acid sequence binding to the glycolipid of the presynaptic membrane and the amino acid sequence specifically binding to the protein present on the presynaptic membrane. On the other hand, endocytosis of type A BoNT (BoNT/A) is a synaptic vesicle glycoprotein 2C (synaptic vesicle glycoprotein) in which A2 peptide (1040-1053, composed of 13 specific amino acids) in RBD exists on the presynaptic membrane. It is known that this is possible by specifically binding to the luminal domain of 2C, SV2C), and the binding is expected to be stronger than the binding of other (BG) types of BoNT to presynaptic proteins.
세포 내 기능을 보일 것이라 기대되는 특정 고분자물질, 즉, 활성 물질(active material)은 시냅스 전막에 부착되어 세포 내로 함입되어 흡수될 때는 시냅스 전막으로부터 소포를 형성하여 흡수된다. 해당 고분자가 세포 내 기능을 나타내기 위해서는 소포 내강으로부터 세포질로 탈출하는 과정(소포탈출)이 필요하다. 한편, 소포를 탈출하여 세포질로의 이동을 돕는, 바이러스, 박테리아, 동물, 식물 등으로부터 자연유래되거나 또는 때때로 기능성을 향상시키기 위해 아미노산 시퀀스를 합성/변형한 다양한 펩티드들(endosomal escape peptides, EEPs)이 많은 연구들을 통해 알려져 있다(일반적으로 30개 이하의 아미노산).Specific high molecular substances expected to exhibit intracellular functions, ie, active materials, are absorbed by forming vesicles from the presynaptic membrane when they are attached to the presynaptic membrane and are absorbed and absorbed into the cell. In order for the polymer to exhibit intracellular functions, a process of escaping from the vesicle lumen to the cytoplasm (vesicle escape) is required. On the other hand, various peptides (endosomal escape peptides, EEPs) that are naturally derived from viruses, bacteria, animals, plants, etc., which help to escape vesicles and move to the cytoplasm, or sometimes synthesize/modify amino acid sequences to improve functionality It is known from many studies (usually less than 30 amino acids).
한편, 알츠하이머, 보툴리누스 중독증, 통증, 다한증, 피부주름 등 신경계통 이상으로 인해 발생하는 다양한 병증/질환에 대한 다양한 치료법이 존재하지만, 기존의 치료법에서 사용되고 있는 많은 약물/물질들은 신경세포에서만 특이적으로 작용하기 보다는 광역적인 투여를 통해 치료효과를 기대하는 것이 대부분이다. 또한 의료/과학분야에서는 신경세포만을 식별하여 표시하는 기술이 요구되고 있지만, 신경세포만을 특이적으로 식별할 수 있는 기존의 기술은 많지 않고 그 활용이 제한적이다. 한편 단백질 의약품 분자 크기의 증가는 체내의 항원·항체 반응을 일으킬 가능성을 높이기 때문에 치료효과를 감소시킬 가능성이 매우 높을 뿐만 아니라, 피부 층을 통한 치료법에서는 분자 크기가 낮을수록 흡수율이 증대되므로 치료용 약물의 분자 크기를 감소시켜야 할 필요성이 제기된다. 따라서 신경세포를 특이적으로 인식하여 해당 세포의 세포질로 이동할 수 있으면서도 분자량이 작은, 단백질 등의 물질이 의료/과학계에서 필히 요구되나 이에 관한 결과물의 발표는 미흡한 실정이다.On the other hand, although there are various treatments for various diseases/diseases caused by nervous system abnormalities such as Alzheimer's disease, botulism, pain, hyperhidrosis, and skin wrinkles, many drugs/substances used in existing treatments are specific to nerve cells only. Rather than acting, most expect a therapeutic effect through broad-spectrum administration. Also, in the medical/science field, technology for identifying and displaying only nerve cells is required, but there are not many existing technologies that can specifically identify only nerve cells and their application is limited. On the other hand, an increase in the molecular size of a protein drug increases the likelihood of causing an antigen/antibody reaction in the body, so it is highly likely to reduce the therapeutic effect. In the case of treatment through the skin layer, the lower the molecular size, the higher the absorption rate. There is a need to reduce the molecular size of Therefore, materials such as proteins, which can specifically recognize nerve cells and move to the cytoplasm of the cells, are required in the medical/science world, but the publication of the results is insufficient.
본 발명자들은 신경세포를 특이적으로 인식하면서 해당 세포질로 효과적으로 이동시킬 수 있는 작은 분자 크기의 단백질을 개발하기 위하여 예의 노력한 결과, BoNT의 RBD 또는 BoNT/A의 A2 펩티드와 다양한 저분자 EEP들을 도입함으로써 종래의 문제점을 개선함과 동시에 신경세포뿐만 아니라 생체 내에서도 기존의 치료법에 준하는 효능을 띈다는 것을 확인하여 본 발명을 완성하였다.The present inventors made diligent efforts to develop a protein of a small molecular size that can efficiently move into the cytoplasm while specifically recognizing neurons. As a result, by introducing BoNT RBD or BoNT/A A2 peptide and various small molecule EEPs, The present invention was completed by improving the problems of and at the same time confirming that it exhibits efficacy comparable to that of conventional treatment in not only nerve cells but also in vivo.
이에, 본 발명의 목적은, Neuro-active material과 EEP, BoNT/A-A2를 하나 이상 포함함으로써 신경세포를 특이적 인식하고 소포 탈출이 가능한 조성물을 제공하는 것이다. 상기 조성물에는 하기 구조식 1로 표시되는 물질, 구조식 2로 표시되는 물질, 및 구조식 3으로 표시되는 물질로 이루어진 군으로부터 선택되는 물질을 포함될 수 있으나 순서는 무관하다. Accordingly, an object of the present invention is to provide a composition capable of specifically recognizing neurons and vesicle escape by including at least one Neuro-active material, EEP, and BoNT/AA 2 . The composition may include a material selected from the group consisting of a material represented by
[구조식 1][Structural Formula 1]
N-terminal C-terminalN-terminal C-terminal
(Neuro-active material)(EEP1).....(EEPn)(BoNT-RBD 또는 BoNT/A-A2)(Neuro-active material)(EEP 1 ).....(EEP n )(BoNT-RBD or BoNT/AA 2 )
[구조식 2][Structural Formula 2]
N-terminal C-terminalN-terminal C-terminal
(EEP1).....(EEPn)(Neuro-active material)(BoNT-RBD 또는 BoNT/A-A2)(EEP 1 ).....(EEP n ) (Neuro-active material) (BoNT-RBD or BoNT/AA 2 )
[구조식 3][Structural Formula 3]
N-terminal C-terminalN-terminal C-terminal
(EEP1)...(EEPn1)(Neuro-active material)(EEP'1)...(EEP'n2)(BoNT-RBD 또는 BoNT/A-A2)(EEP 1 )...(EEP n1 )(Neuro-active material)(EEP' 1 )...(EEP' n2 )(BoNT-RBD or BoNT/AA 2 )
Neuro-active material : 신경세포에서 작용하도록 유도된 물질Neuro-active material: A substance that is induced to act on nerve cells.
EEP : 엔도소말 이스케이프 펩타이드(endosomal escape peptide)EEP: endosomal escape peptide
BoNT-RBD : 보툴리눔 신경독소의 리셉터 바인딩 도메인(receptor binding domain of botulinum neurotoxin type A)BoNT-RBD: receptor binding domain of botulinum neurotoxin type A
BoNT/A-A2 : 보툴리눔 신경독소 타입A의 A2 펩타이드(A2 peptide of botulinum neurotoxin type A)BoNT / AA 2: botulinum nerve peptide A 2 (A 2 peptide of botulinum neurotoxin type A) of a toxin type A
n, n1, n2는 각각 독립적으로 1 이상의 수이며, EEP는 각각 독립적으로 한번 이상 반복될 수 있다. 즉, [구조식 1]과 [구조식 2]에서 임의의 수 i, j(1≤i≤n; 1≤j≤n)에 대하여 EEPi, EEPj는 같거나 다를 수 있으며, [구조식 3]에서 임의의 수 i, j(1≤i≤n1 or n2; 1≤j≤n1 or n2)에 대하여 EEPi, EEPj, EEP'i, EEP'j는 같거나 다를 수 있다.n, n 1 , and n 2 are each independently a number of 1 or more, and EEP may be independently repeated one or more times. That is, EEPi and EEPj may be the same or different for arbitrary numbers i and j (1≤i≤n; 1≤j≤n) in [Formula 1] and [Formula 2], and in [Formula 3], For numbers i and j (1≤i≤n 1 or n 2 ; 1≤j≤n 1 or n 2 ), EEPi, EEPj, EEP'i, and EEP'j may be the same or different.
본 발명의 다른 목적은, 상기 뉴런을 특이적 인식하고 소포 탈출이 가능한 조성물을 유효성분으로 포함하는, 신경병증 예방/치료 또는 표시용 약학적 조성물을 제공하는 것이다.Another object of the present invention is to provide a pharmaceutical composition for preventing/treating or displaying neuropathy, comprising a composition capable of specifically recognizing the neurons and vesicle escape as an active ingredient.
본 발명의 또 다른 목적은, 상기 뉴런을 특이적 인식하고 소포 탈출이 가능한 조성물을 유효성분으로 포함하는, 화장료 조성물을 제공하는 것이다.Another object of the present invention is to provide a cosmetic composition comprising a composition capable of specifically recognizing the neurons and vesicle escape as an active ingredient.
그러나, 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다.However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.
본 발명은 신경-활성 물질(neuro-active material)에 보툴리눔 독소 단백질의 리셉터 바인딩 도메인(receptor binding domain) 또는 보툴리눔 독소 타입A의 A2 펩타이드(A2 peptide of botulinum neurotoxin type A)가 결합된 신경세포 표적용 단백질을 제공한다.The present invention relates to a neuronal target in which a receptor binding domain of botulinum toxin protein or A2 peptide of botulinum neurotoxin type A is bound to a neuro-active material provide protein.
또한, 본 발명은 신경-활성 물질(neuro-active material)에 보툴리눔 독소 단백질의 리셉터 바인딩 도메인(receptor binding domain) 또는 보툴리눔 독소 타입A의 A2 펩타이드(A2 peptide of botulinum neurotoxin type A)가 결합된 신경세포 표적용 단백질을 유효성분으로 포함하는 신경병증의 예방 또는 치료용 약학적 조성물을 제공한다. In addition, the present invention relates to a neuronal cell in which a receptor binding domain of botulinum toxin protein or A2 peptide of botulinum neurotoxin type A is bound to a neuro-active material. It provides a pharmaceutical composition for preventing or treating neuropathy comprising a target protein as an active ingredient.
본 발명의 일 구체예에 있어서, 상기 약학적 조성물에 포함되어 있는 신경세포 표적용 단백질의 신경-활성 물질은 신경병증 치료용 물질이며, 바람직하게는 보툴리눔 신경독소 경쇄, 저분자 신경-활성 물질, 저분자 신경-활성 물질을 함유하는 담체, 또는 이의 조합 등일 수 있으나, 신경병증을 치료하는데 사용되고 있는 물질이라면 이에 제한되지 않는다.In one embodiment of the present invention, the neuro-active substance of the neuron-targeting protein contained in the pharmaceutical composition is a substance for treating neuropathy, preferably botulinum neurotoxin light chain, a small molecule neuro-active substance, a small molecule It may be a carrier containing a neuro-active substance, or a combination thereof, but is not limited thereto as long as it is a substance used to treat neuropathy.
본 발명의 다른 구체예에 있어서, 상기 신경병증(neuropathy)은 바람직하게는 사시, 안검경련, 성대장애, 사경, 심근장애, 궤양, 위산역류질환, 식욕감소, 췌장질환, 튼 살, 절박성 요실금, 치열, 소아마비, 근육통, 엉덩이 기형, 다한증, 허리통증, 경부통, 만성두통, 뇌신경장애 등이나, 말초신경계의 기능적 장애 및 병적 변화를 나타내는 질병이라면 이에 제한되지 않는다.In another embodiment of the present invention, the neuropathy is preferably strabismus, blepharospasm, vocal cord disorder, torticollis, myocardial disorder, ulcer, acid reflux disease, decreased appetite, pancreatic disease, stretch marks, urge incontinence, Fever, polio, myalgia, hip deformity, hyperhidrosis, back pain, neck pain, chronic headache, cranial nerve disorder, etc., but not limited to diseases that show functional disorders and pathological changes in the peripheral nervous system.
또한, 본 발명은 신경-활성 물질(neuro-active material)에 보툴리눔 독소 단백질의 리셉터 바인딩 도메인(receptor binding domain) 또는 보툴리눔 독소 타입A의 A2 펩타이드(A2 peptide of botulinum neurotoxin type A)가 결합된 신경세포 표적용 단백질을 유효성분으로 포함하는 화장료 조성물을 제공한다.In addition, the present invention relates to a neuronal cell in which a receptor binding domain of botulinum toxin protein or A2 peptide of botulinum neurotoxin type A is bound to a neuro-active material. It provides a cosmetic composition comprising a target protein as an active ingredient.
본 발명의 일 구체예에 있어서, 상기 화장료 조성물에 포함되어 있는 신경세포 표적용 단백질의 신경-활성 물질은 바람직하게는 주름 방지를 위해 사용되는 물질일 수 있으며, 더욱 바람직하게는 보툴리눔 신경독소 경쇄, 저분자 신경-활성 물질, 저분자 신경-활성 물질을 함유하는 담체, 또는 이의 조합 등일 수 있으나, 미용 용도로 사용되고 있는 물질이라면 이에 제한되지 않는다.In one embodiment of the present invention, the neuro-active material of the protein for nerve cell targeting contained in the cosmetic composition may be a material used for preventing wrinkles, and more preferably, a botulinum neurotoxin light chain; It may be a low-molecular neuroactive substance, a carrier containing a low-molecular neuro-active substance, or a combination thereof, but is not limited thereto as long as it is a substance used for cosmetic purposes.
또한, 본 발명의 신경-활성 물질(neuro-active material)에 보툴리눔 독소 단백질의 리셉터 바인딩 도메인(receptor binding domain) 또는 보툴리눔 독소 타입A의 A2 펩타이드(A2 peptide of botulinum neurotoxin type A)가 결합된 신경세포 표적용 단백질을 유효성분으로 포함하는 신경계 뉴런 표시용 조성물을 제공한다.In addition, a neuronal cell in which a receptor binding domain of botulinum toxin protein or A2 peptide of botulinum neurotoxin type A is bound to the neuro-active material of the present invention It provides a composition for displaying a nervous system neuron comprising a target protein as an active ingredient.
본 발명의 일 구체예에 있어서, 상기 신경계 뉴런 표시용 조성물에 포함되어 있는 신경세포 표적용 단백질의 신경-활성 물질은 바람직하게는 뉴런을 선택적으로 발색시키는 물질일 수 있으며, 더욱 바람직하게는 형광 단백질, 저분자 형광 화합물, 저분자 형광 화합물을 함유하는 담체, 미세 반도체 입자, 또는 이의 조합 등일 수 있으나, 뉴런을 관찰할 수 있도록 하는 물질이라면 이에 제한되지 않는다.In one embodiment of the present invention, the neuronal-active material of the neuron-targeting protein included in the composition for displaying neurons in the nervous system may be a material that selectively develops neurons, more preferably a fluorescent protein. , a low-molecular-weight fluorescent compound, a carrier containing a low-molecular fluorescent compound, fine semiconductor particles, or a combination thereof, but is not limited thereto as long as it is a material that enables observation of neurons.
본 발명의 다른 구체예에 있어서, 상기 신경세포 표적용 단백질은 엔도소말 이스케이프 펩타이드(endosomal escape peptide)가 하나 이상 추가로 결합된 형태일 수 있으며, 상기 엔도소말 이스케이프 펩타이드는 바람직하게는 서열번호 76 내지 98으로 이루어진 군으로부터 선택되는 어느 하나 이상의 아미노산 서열을 포함할 수 있다. 또한, 상기 엔도소말 이스케이프 펩타이드가 결합된 형태에는 제한이 없다.In another embodiment of the present invention, the protein for targeting nerve cells may be in a form in which one or more endosomal escape peptides are additionally bound, and the endosomal escape peptide is preferably SEQ ID NO: 76 to It may include any one or more amino acid sequences selected from the group consisting of 98. In addition, there is no limitation in the form in which the endosomal escape peptide is bound.
본 발명의 또 다른 구체예에 있어, 상기 보툴리눔 독소 단백질의 리셉터 바인딩 도메인은 바람직하게는 서열번호 99 내지 115으로 이루어진 군으로부터 선택되는 어느 하나 이상의 아미노산 서열을 포함하는 것을 특징으로 한다.In another embodiment of the present invention, the receptor binding domain of the botulinum toxin protein preferably comprises any one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 99 to 115.
본 발명의 또 다른 구체예에 있어서, 상기 보툴리눔 독소 타입A의 A2 펩타이드는 바람직하게는 서열번호 116 내지 120으로 이루어진 군으로부터 선택되는 어느 하나 이상의 아미노산 서열을 포함하는 것을 특징으로 한다. 본 발명의 또 다른 구체예에 있어서, 상기 신경-활성 물질은 바람직하게는 보툴리눔 신경독소 경쇄이며, 상기 보툴리눔 신경독소 경쇄는 서열번호 1 내지 70으로 이루어진 군으로부터 선택되는 하나 이상의 아미노산 서열을 포함하는 것을 특징으로 한다.In another embodiment of the present invention, the botulinum toxin type A A2 peptide preferably comprises any one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 116 to 120. In another embodiment of the present invention, the neuro-active substance is preferably a botulinum neurotoxin light chain, and the botulinum neurotoxin light chain comprises at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 70 characterized.
본 발명의 또 다른 구체예에 있어서, 상기 저분자 신경-활성 물질은 플라보노이드(flavonoid), 스테로이드(sterioid), 알칼로이드(alkaloid), 스틸베노이드(stilbenoid), 찰코노이드(chalconoid), 컬큐민(curcumin), 베라파밀(verapamil), 6-쇼가올(6-shogaol), 브로모데옥 시우리딘(bromodeoxyuridine), 5-플루오로우라신(5-fluorouracil), 요오드데옥시우리딘(iododeoxyuridine), 도파민(dopamine), 노르에피네프린(norepinephrine), 레티노산(retinoic acid), 파클리탁셀(paclitaxel), 카르보플라틴(carboplatin), 아이소나이아지드(isoniazid), 레보도파(levodopa), 카르무스틴(carmustine) 등일 수 있으며, 상기 플라보노이드에는 Quercetin, Fisetin, Myricetin, Luteolin, Laricitrin, Syringetin, Apigenin, Amontoflavone, Catechin 등이 포함될 수 있으며, 상기 스테로이드에는 Tetrahydrodeoxycorticosterone, Androstane, 3α-Androstanediol, Cholestane, Eltanolone, Brexanolone, Pregnenolone sulfate, Epipregnanolone, Ganaxolone, Isopregnanolone, Dehydroepiandrosterone, Dehydroepiandrosterone sulfate, 24(S)-Hydroxychlesterol, Pregnenolone, Progesterone, Estradiol, Corticosterone, Allopregnanolone, Aloradine, 3β-Metholxypregnenolone 등이 포함될 수 있으며, 상기 알칼로이드에는 Tryptoline, Mellpaladine, β-Carboline 등이 포함될 수 있으며, 상기 스틸베노이드에는 Resveratrol, Piceatannol 등이 포함될 수 있으며, 상기 찰코노이드에는 Butein이 포함될 수 있다.In another embodiment of the present invention, the small molecule neuro-active substance is a flavonoid, a steroid, an alkaloid, a stilbenoid, a chalconoid, and a curcumin. , verapamil, 6-shogaol, bromodeoxyuridine, 5-fluorouracil, iododeoxyuridine, dopamine , norepinephrine, retinoic acid, paclitaxel, carboplatin, isoniazid, levodopa, carmustine, etc. may be Flavonoids may include Quercetin, Fisetin, Myricetin, Luteolin, Laricitrin, Syringetin, Apigenin, Amontoflavone, Catechin, and the like, and the steroids include Tetrahydrodeoxycorticosterone, Androstane, 3α-Androstanediol, Cholestane, Eltanolone, Pregnopenol sulfate, Eltanolone, Brexanolone, anapregnanolone, Brexanolone, etc. , Dehydroepiandrosterone, Dehydroepiandrosterone sulfate, 24(S)-Hydroxychlesterol, Pregnenolone, Progesterone, Estradiol, Corticosterone, Allopregnanolone, Aloradine, 3β-Metholxypregnenolone, etc. may be included, and the alkaloids may include Tryptoline, β-Carboline, etc. , The stilbenoid may include Resveratrol, Piceatannol, and the like, and the chalconoid may include Butein.
본 발명의 또 다른 구체예에 있어서, 상기 형광 단백질은 바람직하게는 서열번호 71 내지 75으로 이루어진 군으로부터 선택되는 아미노산 서열을 하나 이상 포함할 수 있다.In another embodiment of the present invention, the fluorescent protein may include one or more amino acid sequences preferably selected from the group consisting of SEQ ID NOs: 71 to 75.
본 발명의 또 다른 구체예에 있어서, 상기 저분자 형광 화합물은 크산틴 유도체(xanthene derivatives), 시아닌 유도체(cyanine derivatives), 스쿠아레인 유도체(Squaraine derivatives), 스쿠아레인 로탁산 유도체(Squaraine Rotaxane derivatives), 나프탈렌 유도체(naphthalene derivatives), 쿠마린 유도체(coumarin derivatives), 옥사디아졸 유도체(oxadiazole derivatives), 안트라센 유도체(anthracene derivatives), 피렌 유도체(pyrene derivatives), 옥사진 유도체(oxazine derivatives), 아크리딘 유도체(acridine derivatives), 아릴메틴 유도체(arylmethine derivatives), 테트라피롤 유도체(tetrapyrrole derivatives) 등 일 수 있으며, 상기 유도체는 화합물의 일부를 변형시켜 얻어지는 유사한 화합물을 총칭하는 말로서, 크산틴 유도체에는 Fluorescein, Rhodamine, Oregon green, Eosin, Texas red 등이 포함될 수 있으며, 시아닌 유도체에는 Cyanine Indocarbocyanine, Oxacarbocyanine, Thiacarbocyanine, merocyanine 등이 포함될 수 있으며, 스쿠아레인 유도체에는 Seta dyes, Square dyes 등이 포함될 수 있으며, 스쿠아레인 로탁산 유도체에는 SeTau dyes 등이 포함될 수 있으며, 나프탈렌 유도체에는 Densyl/prodan derivatives 등이 포함될 수 있으며, 옥사디아졸 유도체에는 Pyridyloxazole, Nitrobenzoxadiazole, Benzoxadiazole 등이 포함될 수 있으며, 안트라센 유도체에는 Anthraquinones, DRAQ5, DRAQ7, CyTRAK Orange 등이 포함될 수 있으며, 피렌 유도체에는 Cascade blue 등이 포함될 수 있으며, 옥사진 유도체에는 Nile red, Nile blue, Cresyl violet, Oxazine 170 등이 포함될 수 있으며, 아크리딘 유도체에는 Proflavin, Acridine orange, Acridine yellow 등이 포함될 수 있으며, 아릴메틴 유도체에는 Auramine, Crystal violet, Malachite green 등이 포함될 수 있으나, 저분자 형광 화합물의 유도체라면 이에 제한되지 않는다.In another embodiment of the present invention, the low molecular weight fluorescent compound is xanthene derivatives, cyanine derivatives, squaraine derivatives, and squaraine rotaxane derivatives. , naphthalene derivatives, coumarin derivatives, oxadiazole derivatives, anthracene derivatives, pyrene derivatives, oxazine derivatives, acridine derivatives (acridine derivatives), arylmethine derivatives, tetrapyrrole derivatives, etc., and the derivative is a generic term for a similar compound obtained by modifying a part of the compound. Xanthine derivatives include Fluorescein, Rhodamine, Oregon green, Eosin, Texas red, etc. may be included, and the cyanine derivative may include Cyanine Indocarbocyanine, Oxacarbocyanine, Thiacarbocyanine, merocyanine, etc., and the squaraine derivative may include Seta dyes, Square dyes, etc. Taxane derivatives may include SeTau dyes, etc., naphthalene derivatives may include Densyl/prodan derivatives, etc., oxadiazole derivatives may include Pyridyloxazole, Nitrobenzoxadiazole, Benzoxadiazole, etc., and anthracene derivatives include Anthraquinones, DRAQ5, DRAQ7, CyTRAK Orange, etc. may be included, pyrene oil The conductor may include Cascade blue, etc., and the oxazine derivatives may include Nile red, Nile blue, Cresyl violet, Oxazine 170, etc., and the acridine derivatives may include Proflavin, Acridine orange, Acridine yellow, etc., and aryl The methine derivative may include auramine, crystal violet, malachite green, and the like, but is not limited thereto as long as it is a derivative of a low molecular weight fluorescent compound.
본 발명의 또 다른 구체예에 있어서, 상기 미세 반도체 입자는 바람직하게는 퀀텀닷(quantum dot)이나, 세포 내에서 세포를 관찰할 수 있도록 하는 표지 물질이라면 제한이 없다.In another embodiment of the present invention, the micro semiconductor particles are preferably quantum dots, but there is no limitation as long as it is a labeling material that allows the observation of cells in cells.
본 발명의 또 다른 구체예에 있어서, 상기 담체는 바람직하게는 유기미세입자, 무기미세입자, 자가조립구조체(self-assembled structure) 등의 담체(전달) 시스템일 수 있으며, 더욱 바람직하게는 금 나노입자(gold nanoparticle), 철 나노입자(magnetic nanoparticle), 실리카 나노입자(silica nanoparticle), 라텍스 나노입자(latex nanoparticle), 지질 나노입자(lipid nanoparticle), 탄소 나노구조체(carbon nanoplate/nanotube), 리포솜(liposome), 나노디스크(nanodisc) 등 일 수 있으나, 이에 제한되지 않는다.In another embodiment of the present invention, the carrier is preferably a carrier (delivery) system such as organic microparticles, inorganic microparticles, and self-assembled structures, and more preferably gold nano-particles. Gold nanoparticles, magnetic nanoparticles, silica nanoparticles, latex nanoparticles, lipid nanoparticles, carbon nanoplates/nanotubes, liposomes ( liposome), nanodisc, etc., but is not limited thereto.
또한, 본 발명은 상기 신경세포 표적용 단백질을 유효성분으로 포함하는 조성물을 개체에 투여하는 단계를 포함하는 신경병증의 예방, 개선 또는 치료 방법을 제공한다.In addition, the present invention provides a method for preventing, improving or treating neuropathy, comprising administering to an individual a composition comprising the protein for targeting nerve cells as an active ingredient.
또한, 본 발명은 상기 신경세포 표적용 단백질을 유효성분으로 포함하는 조성물의 신경병증 예방, 개선 또는 치료 용도를 제공한다.In addition, the present invention provides a use for preventing, improving or treating neuropathy of a composition comprising the neuron-targeting protein as an active ingredient.
또한, 본 발명은 상기 신경세포 표적용 단백질의 신경병증 질환에 이용되는 약제를 생산하기 위한 용도를 제공한다.In addition, the present invention provides a use for producing a medicament used in a neuropathic disease of the protein for the nerve cell target.
본 발명의 뉴런을 특이적 인식하고 소포 탈출이 가능한 조성물은 신경계만 특이적으로 식별하여 작용하는 능력을 갖으면서도 신경세포질내 함입이 가능하므로, 신경계 이상으로 유발된 다양한 병증의 치료(경감/개선)용도와 신경세포만을 가시적으로 표시하는 용도로의 사용이 기대된다. 또한 구조를 단순화하여 분자량을 줄인 것으로, 생산과정의 단순화를 도모할 수 있어 경제적이고, 작은 분자량에 의해 체내 항체, 항원 반응으로부터 더 자유로울 수 있으며, 나아가 화장료로 사용되는 경우 분자의 크기가 작아 흡수율이 증대된다는 이점이 있으므로, 관련된 의료사업분야 및 화장품 산업 분야 등에 다양하게 이용될 수 있을 것으로 기대된다.The composition capable of specifically recognizing neurons and vesicle escape of the present invention has the ability to specifically identify and act only on the nervous system and yet can penetrate into the neurocytoplasm, so treatment (relief/improvement) of various conditions induced by nervous system abnormalities It is expected to be used for the purpose of visually displaying only the use and nerve cells. In addition, by reducing the molecular weight by simplifying the structure, it is economical because it can simplify the production process, and it can be more free from antibody and antigen reactions in the body due to the small molecular weight. Furthermore, when used as a cosmetic, the absorption rate is reduced due to the small size of the molecule Since it has the advantage of increasing, it is expected that it can be used in various fields such as related medical business fields and cosmetic industries.
도 1은 본 발명에서의 뉴런을 특이적 인식하고 소포 탈출이 가능한 조성물의 일례로서 재조합 보툴리눔 신경독소가 뉴런 내로 흡수되고, 세포질로 이동한 후 SNARE 복합체 형성을 저해하는 과정을 나타낸 모식도이다.
도 2a 및 2b는 BoNTs LC, EEP, 및 BoNT/A-A2 단백질의 가능한 결합구조를 예시한 것이다.
도 3은 BoNTs LC를 생산하는데 사용된 클로닝 벡터 맵의 구조를 나타낸 것이다.
도 4는 경쟁적 엘라이자(ELAISA) 기법을 사용하여 노르에피네프린 농도를 측정하는 실험방법을 나타낸 것이다.
도 5는 재조합 보툴리눔 신경독소가 분화된 PC12 세포에서 노르아드레날린 방출량에 미치는 영향을 실험한 결과이다.
도 6은 재조합 보툴리눔 신경독소의 국소 근육약화효능을 확인하기 위해 마
우스를 대상으로 한 디지트 어브덕션 스코어(digit abduction score; DAS) 어세이(assay) 결과이다.
도 7은 본 발명에서의 뉴런을 특이적 인식하는 조성물의 일례로서의 녹색형광단백질(enhanced green fluorescent protein, EGFP)을 포함한 뉴런 표시용 조성물들을 생산하기 위한 유전자의 모식도(도 7B)와 peptide A2' 또는 peptide A2에 결합하는 수용체(synaptic vesicle glycoprotein 2C, SV2C)를 생산하기 위한 유전자의 모식도(도 7C)를 나타낸 것이다(도 7A, peptide A2'와 peptide A2의 아미노산 서열).
도 8은 도 7의 EGFP를 포함하는 뉴런 표시용 조성물을 생산하는데 사용된 클로닝 벡터 맵(도 8A)과 SV2C를 생산하는데 사용된 클로닝 벡터 맵(도 8B)의 구조를 나타낸 것이다.
도 9는 도 7의 EGFP를 포함하는 뉴런 표시용 조성물과 SV2C을 발현 및 정제 후의 SDS-PAGE젤 전기영동 결과이다.
도 10은 도 7의 EGFP를 포함하는 뉴런 표시용 조성물과, SV2C가 결합되어 있는 bead와의 반응 후의 반응물의 SDS-PAGE젤 전기영동 결과이다.
도 11은 도 7의 EGFP를 포함하는 뉴런 표시용 조성물을 처리한 분화된 PC12 세포의 공초점 레이저 형광 현미경 사진이다.1 is a schematic diagram showing a process of inhibiting SNARE complex formation after recombinant botulinum neurotoxin is absorbed into neurons and migrates to the cytoplasm as an example of a composition capable of specifically recognizing neurons and vesicle escape in the present invention.
2a and 2b illustrate possible binding structures of BoNTs LC, EEP, and BoNT/A-A2 proteins.
3 shows the structure of the cloning vector map used to produce BoNTs LCs.
Figure 4 shows an experimental method for measuring the concentration of norepinephrine using a competitive ELISA (ELAISA) technique.
5 is a result of an experiment on the effect of recombinant botulinum neurotoxin on the amount of noradrenaline released in differentiated PC12 cells.
6 is a diagram to confirm the local muscle-weakening effect of recombinant botulinum neurotoxin.
It is a digit abduction score (DAS) assay result for mice.
7 is a schematic diagram of a gene for producing compositions for displaying neurons including enhanced green fluorescent protein (EGFP) as an example of a composition for specifically recognizing neurons in the present invention ( FIG. 7B ) and peptide A2' or A schematic diagram (FIG. 7C) of a gene for producing a receptor (synaptic vesicle glycoprotein 2C, SV2C) that binds to peptide A2 is shown (FIG. 7A, amino acid sequences of peptide A2' and peptide A2).
8 shows the structures of the cloning vector map (FIG. 8A) used to produce the composition for displaying neurons including EGFP of FIG. 7 and the cloning vector map (FIG. 8B) used to produce SV2C.
9 is an SDS-PAGE gel electrophoresis result after expression and purification of the neuron-displaying composition and SV2C containing the EGFP of FIG. 7 .
10 is an SDS-PAGE gel electrophoresis result of the reaction product after reaction between the neuron-displaying composition including the EGFP of FIG. 7 and the bead to which SV2C is bound.
FIG. 11 is a confocal laser fluorescence micrograph of differentiated PC12 cells treated with the neuron-displaying composition of FIG. 7 .
본 발명자들은 뉴런을 특이적 인식하고 소포 탈출이 가능한 조성물을 개발하기 위해 노력한 결과, 기존 BoNT(botulinum neurotoxin) 단백질의 RBD(receptor binding domain) 또는 BoNT/A의 A2 펩티드와 뉴런 내 소포 탈출이 가능토록 다양한 저분자 EEP(endosomal escape peptide)들을 신경-활성 물질에 도입함으로써 신경계만 특이적으로 식별하여 작용하는 능력을 가지면서도 생체 신경계를 대상으로 우수한 효과를 나타낸다는 것을 확인하여 본 발명을 완성하였다.As a result of the present inventors' efforts to develop a composition capable of specifically recognizing neurons and vesicle escape, existing BoNT (botulinum neurotoxin) protein RBD (receptor binding domain) or BoNT / A A2 peptide and vesicle escape in neurons are possible By introducing various low-molecular endosomal escape peptides (EEPs) into nerve-active substances, the present invention was completed by confirming that they exhibit excellent effects on the biological nervous system while having the ability to specifically identify and act only on the nervous system.
본 발명의 일 실시예에서는, SNARE(Soluble Nethylameimide-sensitive factor Attachment Protein Receptor; SNAP Receptor) 복합체 형성 저해용 재조합 보툴리눔 신경독소를 생산하기 위한 플라스미드의 제조방법 및 상기 플라스미드를 이용한 재조합 보툴리눔 신경독소 생산방법을 확인하였다(실시예 1 및 2 참조).In one embodiment of the present invention, a method for producing a plasmid for producing a recombinant botulinum neurotoxin for inhibiting the formation of a SNARE (Soluble Nethylameimide-sensitive factor Attachment Protein Receptor; SNAP Receptor) complex and a method for producing a recombinant botulinum neurotoxin using the plasmid are provided. was confirmed (see Examples 1 and 2).
본 발명의 다른 실시예에서는, 분화된 PC12 세포를 이용하여 노르에피네프린 방출량을 분석한 결과 재조합 보툴리눔 신경독소가 노르에피네프린 방출을 억제하는 효과가 있음을 확인하였다(실시예 3 참조).In another example of the present invention, as a result of analyzing the amount of norepinephrine released using differentiated PC12 cells, it was confirmed that the recombinant botulinum neurotoxin had the effect of inhibiting the release of norepinephrine (see Example 3).
본 발명의 또 다른 실시예에서는, 마우스를 이용하여 디지트 어브덕션 스코어(digit abduction score; DAS) 어세이(assay)를 수행한 결과 재조합 보툴리눔 신경독소가 근육약화효능을 가지고 있음을 확인하였다(실시예 4 참조).In another embodiment of the present invention, as a result of performing a digit abduction score (DAS) assay using a mouse, it was confirmed that the recombinant botulinum neurotoxin has a muscle-weakening effect (Example) see 4).
본 발명의 다른 실시예에서는, 뉴런 특이적 표시 기능을 지닌 녹색형광단백질을 생산하기 위한 플라스미드의 제조방법 및 상기 플라스미드를 이용한 뉴런 표시용 재조합 녹색형광단백질의 생산방법을 확인하였다(실시예 5 및 6 참조).In another embodiment of the present invention, a method for producing a plasmid for producing a green fluorescent protein having a neuron-specific display function and a method for producing a recombinant green fluorescent protein for displaying a neuron using the plasmid were confirmed (Examples 5 and 6) Reference).
본 발명의 다른 실시예에서는, 제조된 뉴런 표시용 재조합 녹색형광단백질과 뉴런 세포막에 존재하는 SV2C(synaptic vesicle glycoprotein 2C) 사이에 결합력이 있는 것을 확인하였다(실시예 7 참조).In another embodiment of the present invention, it was confirmed that there is a binding force between the prepared recombinant green fluorescent protein for neuronal display and synaptic vesicle glycoprotein 2C (SV2C) present in the neuronal cell membrane (see Example 7).
본 발명의 또 다른 실시예에서는, 분화된 PC12 세포와 공초점 레이저 형광 현미경(confocal laser fluorescence microscope, CLFM)을 이용하여 얻은 이미지의 결과 뉴런 표시용 재조합 녹색형광단백질이 상기 세포에 잘 부착되고 세포 내로 잘 흡수되는 것을 확인하였다(실시예 8 참조).In another embodiment of the present invention, as a result of images obtained using differentiated PC12 cells and a confocal laser fluorescence microscope (CLFM), the recombinant green fluorescent protein for neuron display is well attached to the cells and enters the cells. It was confirmed that it was well absorbed (see Example 8).
이에, 본 발명은 하기 구조식 1로 표시되는 물질, 구조식 2로 표시되는 물질, 및 구조식 3으로 표시되는 물질로 이루어진 군으로부터 선택되는 하나 이상의 단백질을 포함하는, 뉴런을 특이적 인식하고 소포 탈출이 가능한 조성물을 제공한다.Accordingly, the present invention includes at least one protein selected from the group consisting of a material represented by the following
[구조식 1][Structural Formula 1]
N-terminal C-terminalN-terminal C-terminal
(Neuro-active material)(EEP1).....(EEPn)(BoNT-RBD 또는 BoNT/A-A2)(Neuro-active material)(EEP 1 ).....(EEP n )(BoNT-RBD or BoNT/AA 2 )
[구조식 2][Structural Formula 2]
N-terminal C-terminalN-terminal C-terminal
(EEP1).....(EEPn)(Neuro-active material)(BoNT-RBD 또는 BoNT/A-A2)(EEP 1 ).....(EEP n ) (Neuro-active material) (BoNT-RBD or BoNT/AA 2 )
[구조식 3][Structural Formula 3]
N-terminal C-terminalN-terminal C-terminal
(EEP1)...(EEPn1)(Neuro-active material)(EEP'1)...(EEP'n2)(BoNT-RBD 또는 BoNT/A-A2)(EEP 1 )...(EEP n1 )(Neuro-active material)(EEP' 1 )...(EEP' n2 )(BoNT-RBD or BoNT/AA 2 )
Neuro-active material : 신경-활성 물질, 신경세포에서 작용하도록 유도된 물질, 신경세포에서 활성을 나타내는 단백질Neuro-active material: A neuro-active substance, a substance induced to act on neurons, and a protein that exhibits activity in neurons
EEP : 엔도소말 이스케이프 펩타이드(endosomal escape peptide)EEP: endosomal escape peptide
BoNT-RBD : 보툴리눔 신경독소의 리셉터 바인딩 도메인(receptor binding domain of botulinum neurotoxin type A)BoNT-RBD: receptor binding domain of botulinum neurotoxin type A
BoNT/A-A2 : 보툴리눔 신경독소 타입A의 A2 펩타이드(A2 peptide of botulinum neurotoxin type A)BoNT / AA 2: botulinum nerve peptide A 2 (A 2 peptide of botulinum neurotoxin type A) of a toxin type A
n, n1, n2는 각각 독립적으로 1 이상의 수이며, EEP는 각각 독립적으로 한번 이상 반복될 수 있다. 즉, [구조식 1]과 [구조식 2]에서 임의의 수 i, j(1≤i≤n; 1≤j≤n)에 대하여 EEPi, EEPj는 같거나 다를 수 있으며, [구조식 3]에서 임의의 수 i, j(1≤i≤n1 or n2; 1≤j≤n1 or n2)에 대하여 EEPi, EEPj, EEP'i, EEP'j는 같거나 다를 수 있다.n, n 1 , and n 2 are each independently a number of 1 or more, and EEP may be independently repeated one or more times. That is, EEPi and EEPj may be the same or different for arbitrary numbers i and j (1≤i≤n; 1≤j≤n) in [Formula 1] and [Formula 2], and in [Formula 3], For numbers i and j (1≤i≤n 1 or n 2 ; 1≤j≤n 1 or n 2 ), EEPi, EEPj, EEP'i, and EEP'j may be the same or different.
본 발명에서 사용하는 용어 "SNARE(Soluble Nethylameimide-sensitive factor Attachment Protein Receptor; SNAP Receptor)" 란 효모 및 포유류 세포에 존재하는 60개 이상의 멤버로 구성된 거대 단백질 수퍼패밀리로, SNARE 단백질의 주된 역할은 소포 융합(vesicle fusion)을 매개하는 것이다. 즉, 리소좀과 같은 기관(compartmets)에 부착된 이들은 표적 막과 소포의 융합을 매개한다. 구체적인 예로서, SNARE는 신경세포에서 시냅스전막(presynaptic membrane)과 시냅스소포(synaptic vesicle)의 결합(docking)에 관여한다.As used herein, the term "Soluble Nethylameimide-sensitive Factor Attachment Protein Receptor (SNAP Receptor)" is a large protein superfamily composed of more than 60 members present in yeast and mammalian cells, and the main role of the SNARE protein is vesicle fusion mediating vesicle fusion. That is, those attached to compartments such as lysosomes mediate the fusion of the vesicle with the target membrane. As a specific example, SNARE is involved in the binding (docking) of a presynaptic membrane and a synaptic vesicle in a neuron.
한편 근육의 이완과 수축을 조절하기 위하여 근육의 상층에는 신경근육 접합부(neuromuscular junction)가 있으며, 이 신경 말단(nerve terminal)에는 시냅스 소포가 장전되어 있다. 근육들은 일종의 신경소포 내부에서 전달되는 신경전달물질의 메시지를 받아 수축하게 되며, 이와 같이 신경전달물질들이 방출되려면, SNARE 단백질들이 복합체를 형성함으로써 신경물질들이 근육과 도킹해야 된다. 구체적으로, 신경전달물질의 배출 시에 신경전달물질을 담고 있는 시냅스 소포는 시냅스전막과 융합되어야 두 경계 간의 통로가 형성될 수 있으며, 이 때 막 융합의 근원적인 힘은 3종의 단백질 복합체로 존재하는 SNARE에 의해 제공된다. 특히, 시냅스 소포와 시냅스전막 간의 융합에 의해 신경전달물질 배출 통로가 열리게 된다. 표적 막(target membrane)에 부착되어 있는, 신탁신(Syntaxin) 1a 단백질과 SNAP-25 단백질의 복합체인 t-SNARE 복합체와, 소포에 부착되어 있는 v-SNARE가 관여하게 되는데, 이들 SNARE 단백질은 꽈배기처럼 꼬여 있게 된다.On the other hand, in order to control the relaxation and contraction of the muscle, there is a neuromuscular junction in the upper layer of the muscle, and the synaptic vesicle is loaded in the nerve terminal. Muscles contract by receiving a message from a neurotransmitter transmitted from within a kind of neurovesicle, and in order for these neurotransmitters to be released, SNARE proteins form a complex and the neurotransmitters must dock with the muscle. Specifically, when the neurotransmitter is discharged, the synaptic vesicle containing the neurotransmitter must be fused with the presynaptic membrane to form a passage between the two boundaries. At this time, the fundamental force of membrane fusion exists as a complex of three types of proteins. provided by SNARE. In particular, the neurotransmitter release pathway is opened by fusion between the synaptic vesicle and the presynaptic membrane. The t-SNARE complex, which is a complex of Syntaxin 1a protein and SNAP-25 protein, attached to the target membrane, and v-SNARE attached to the vesicle are involved. twisted like
상기 막 융합 시에는 당해 분야에 널리 알려져 있는 바와 같이 지질이중층(lipid bilayer)의 재배열이 일어나게 된다. 생체막들은 물리적으로 서로 강하게 밀어내고 있으므로 이들 막은 자발적으로 융합되지 않고 외부에서 강한 힘이 주어져 막들 간의 반발력을 극복하여야 하는데, 이때 막들 간의 반발력을 이겨낼 정도의 강한 힘을 만들어 내는 것이 SNARE 단백질이다. 즉 SNARE 복합체의 형성은 막 간 반발력을 극복하는 힘의 원천이며, 신경전달물질의 배출을 포함하는 세포 외 배출작용(exocytosis)의 핵심 현상인 것이다.When the membrane is fused, rearrangement of the lipid bilayer occurs, as is well known in the art. Since biological membranes physically repel each other strongly, these membranes do not spontaneously fuse, but a strong external force is applied to overcome the repulsive force between the membranes. At this time, it is the SNARE protein that creates a strong enough force to overcome the repulsive force between the membranes. In other words, the formation of the SNARE complex is a source of strength to overcome the transmembrane repulsion, and is a key phenomenon of exocytosis, including the release of neurotransmitters.
일례로, 모공은 얼굴 피부에 전체적으로 고르게 분포하나, 특히 코와 볼 부위는 육안으로 식별할 수 있을 정도로 많고, 모공의 양상은 성별, 유전적, 노화, 여드름, 만성 자외선 노출 등의 내인성과 외인성 요인에 의해 개인차가 있다. 모공이 넓어지는 이유는 피지가 과도하게 분비되거나 피부노화가 시작되면서 모공 벽을 지지하고 있던 콜라겐 섬유와 탄력 섬유가 변성, 감소함에 따라 피부 탄력이 없어지고 쳐지기 때문이다. 털에 붙어있는 근육(입모근)의 수축 또는 이완은 교감신경과 부교감신경의 지배를 받기 때문에, 모공은 신경조절에 의해 축소 또는 확대될 수 있으며 교감신경은 근육에서 약간 떨어져 있으면서 모공 축소 작용을, 부교감 신경은 근육 가까이에 있으면서 모공 확대 작용을 한다. 선택적으로 부교감 신경을 억제하면 보상 작용으로 교감신경이 작용하여 털에 붙어 있는 근육을 수축시켜 모공이 축소된다.For example, the pores are evenly distributed throughout the skin of the face, but in particular, the nose and cheeks are large enough to be identified with the naked eye, and the shape of the pores is determined by intrinsic and extrinsic factors such as gender, genetics, aging, acne, and chronic UV exposure. There are individual differences by The reason for the enlarged pores is that the skin loses elasticity and sags as the collagen fibers and elastic fibers that support the pore walls degenerate and decrease as sebum is secreted excessively or skin aging begins. Since the contraction or relaxation of the muscle attached to the hair (the trapezius muscle) is controlled by the sympathetic nerve and the parasympathetic nerve, the pores can be reduced or enlarged by neuromodulation, and the sympathetic nerve acts slightly away from the muscle to reduce the pores, parasympathetic. Nerves work to enlarge pores while being close to the muscles. If the parasympathetic nerve is selectively suppressed, the sympathetic nerve acts as a compensatory action to contract the muscles attached to the hair, thereby reducing the pores.
반면에, SNARE 접합과 꼬임 과정이 완전히 완료되지 않으면 막 융합이 실패하고 그에 따라 신경전달물질 방출이 이뤄지지 않아 결국 근육의 움직임이 없게 된다. 이 과정은 자주 사용하는 근육에 의하여 생성되는 주름의 생성을 예방하고 미리 만들어진 주름도 개선할 수 있음을 의미한다. 즉, SNARE 형성 억제 효능에 의하여 근육의 운동에 의한 주름의 생성을 억제하고 생성된 주름을 개선할 수 있다.On the other hand, if the SNARE splicing and twisting process is not fully completed, membrane fusion fails, resulting in no neurotransmitter release, resulting in no muscle movement. This process prevents the generation of wrinkles generated by muscles that are used frequently and means that pre-made wrinkles can also be improved. That is, it is possible to suppress the generation of wrinkles due to muscle movement and improve the generated wrinkles by the SNARE formation inhibitory effect.
한편, 땀샘 주위를 감싸고 있는 근세포의 수축은 땀샘을 자극하여 땀이 방출되도록 하는 역할을 하고 있다. 이러한 땀샘 주위 근세포의 수축 또한 신경세포로부터의 신경전달물질 방출로부터 비롯된다. 근세포-신경접합부 상에서 과도한 신경전달물질의 방출은 과다한 땀 분비를 유발하게 되는데, 이러한 병증을 다한증(hyperhidrosis)이라 한다. 따라서 상기 근세포-신경접합부 상에 존재하는 뉴런 내의 SNARE 복합체 형성 저해는 다한증의 병증을 치료 또는 완화할 수 있다.On the other hand, contraction of the muscle cells surrounding the sweat glands stimulates the sweat glands to release sweat. The contraction of myocytes around these glands also results from the release of neurotransmitters from neurons. Excessive release of neurotransmitters on the myocyte-neuronal junction causes excessive sweating, a condition called hyperhidrosis. Therefore, inhibition of SNARE complex formation in neurons present on the myocyte-neuronal junction can treat or alleviate the condition of hyperhidrosis.
상기 SNARE를 표적으로 하는 대표적인 물질로는 보툴리눔 식중독(botulism) 및 파상풍(tetanus)을 유발하는 박테리아 신경독소(bacterial neurotoxin) 등이 있다. 예컨대, 클로스트리디움 보툴리눔(Clostridium botulinum) 유래의 신경독소는 "보툴리눔 신경독소"로 알려진 약물의 주성분으로, 구체적으로, 보툴리눔 신경독소는 그 주성분인 신경독소가 신경세포에 존재하는 SNARE에 특이적으로 작용 후 복합체 형성을 저해하여 막 융합을 억제함으로써 신경전달물질의 방출을 차단한다. 이에 따라 근육의 움직임이나 교감 또는 부교감 신경계를 억제함으로써 상기와 같은 질환의 치료 효과를 나타내는 것으로 알려져 있다.Representative substances that target the SNARE include bacterial neurotoxin that causes botulism and tetanus. For example, Clostridium botulinum-derived neurotoxin is a main component of a drug known as "botulinum neurotoxin", specifically, botulinum neurotoxin is a neurotoxin whose main component is a neurotoxin present in nerve cells. It blocks the release of neurotransmitters by inhibiting membrane fusion by inhibiting complex formation after action. Accordingly, it is known that by inhibiting muscle movement or the sympathetic or parasympathetic nervous system, the therapeutic effect of the above diseases is exhibited.
상기 보툴리눔 신경독소(botulinum neurotoxin, BoNT) 단백질은 크게 약 100kDa의 중쇄(heavy chain; HC)와 신경세포 내에서 SNARE 복합체 형성 단백질을 절단하는 효소활성을 지니는 약 50 kDa의 경쇄(light chain; LC) 두 부분으로 나누어지며, 여기서 HC은 또다시 신경세포를 인식하여 결합하는 기능을 가진 부분(receptor binding domain, RBD)과 LC을 신경세포질로 이동시키는 기능을 가진 도메인(translocation domain)으로 나누어진다. 상기 신경독소 단백질은 단일 사슬로 박테리아에서 방출된 이후, 자체 프로테아제에 의해 두 사슬로 나눠지고 두 사슬은 이황화 결합을 통해 다시 연결되어 약 150 kDa 크기로 존재한다. The botulinum neurotoxin (BoNT) protein is largely about 100 kDa heavy chain (HC) and about 50 kDa light chain (LC) having enzymatic activity for cleaving the SNARE complex-forming protein in neurons. It is divided into two parts, where the HC is further divided into a receptor binding domain (RBD) having a function of recognizing and binding to a neuron, and a domain having a function of moving LC to the neurocytoplasm (translocation domain). After the neurotoxin protein is released from the bacteria as a single chain, it is split into two chains by its own protease, and the two chains are reconnected through disulfide bonds to exist in a size of about 150 kDa.
상기 BoNT 단백질의 RBD는 시냅스 전막의 당지질과 결합하는 아미노산시퀀스와 시냅스 전막 상에 존재하는 단백질과 특이적으로 결합하는 아미노산시퀀스 중 전자만을 포함하거나 모두 포함하고 있다. 전자와 시냅스 전막 당지질과의 결합(해리상수: 약 10-6-10-7 M)은 후자와 특정 시냅스 전막 단백질과의 결합(해리상수: 10-13-10-14 M)에 비해 비 특이적이며 훨씬 약한 결합력을 가진다고 알려져 있다. 한편 A타입의 BoNT의(BoNT/A) 엔도사이토시스(endocytosis)는 RBD 내의 A2 펩티드(1040-1053, 14개의 특정 아미노산으로 구성)가 시냅스 전막 상에 존재하는 시냅스 소포 당단백질 2C(synaptic vesicle glycoprotein 2C, SV2C)의 루미날 도메인(luminal domain)에 특이적으로 결합함으로써 가능하다 알려져 있으며, 상기 결합은 다른(B-G) 타입의 BoNT와 시냅스 전막 단백질과의 결합에 비해 강력할 것으로 예상되고 있다. 이러한 이유 등으로 인해 BoNT/A는 다른 타입의 BoNT들에 비해 신경세포에 더 빨리 들어간다는 것이 보편적 견해이다.The RBD of the BoNT protein contains only electrons or all of the amino acid sequence binding to the glycolipid of the presynaptic membrane and the amino acid sequence specifically binding to the protein present on the presynaptic membrane. The binding of the former with presynaptic glycolipids (dissociation constant: about 10 -6 -10 -7 M) is non-specific compared to the binding of the latter with specific presynaptic proteins (dissociation constant: 10 -13 -10 -14 M) and is known to have a much weaker binding force. On the other hand, endocytosis of type A BoNT (BoNT/A) is a synaptic vesicle glycoprotein 2C (synaptic vesicle glycoprotein) in which A2 peptide (1040-1053, composed of 14 specific amino acids) in RBD exists on the presynaptic membrane. It is known that this is possible by specifically binding to the luminal domain of 2C, SV2C), and the binding is expected to be stronger than the binding of other (BG) types of BoNT to presynaptic proteins. For these reasons, it is a common view that BoNT/A enters nerve cells faster than other types of BoNTs.
BoNT가 시냅스 전막에 부착되어 세포내로 함입되어 흡수될 때는 시냅스 전막으로부터 소포를 형성하여 흡수된다. BoNT가 SNARE 복합체 형성을 저해하기 위해서는 소포 내강으로부터 세포질로 탈출하는 과정이 필요하며 이때 소포로부터 세포질로 BoNT LC을 이동시키는 부분이 HC의 트렌스로케이션 도메인(translocation domain)이다. 소포 내 pH가 감소하면 상기 트렌스로케이션 도메인 내 알파-헬리시즈(α-helices)가 소포 막에 채널을 형성하게 되며, LC이 상기 채널을 통과하여 세포질 방향으로 이동한 후 HC과의 이황화결합이 끊어지면서 세포질 내에서 활성화된 형태로 존재하게 된다. 본 발명에서 제안하는 재조합 보툴리눔 신경독소 형태는 기존 보툴리눔 신경독소와 유사한 활성에 전제하므로, 상기 BoNT의 트렌스로케이션 도메인(translocation domain)과 유사한 기능(소포탈출)을 갖는 부분이 필요하다. 소포를 탈출하여 세포질로의 이동을 돕는 다양한 펩티드들(endosomal escape peptides, EEPs)은 주로 바이러스, 박테리아, 동물, 식물 등 자연유래로 알려져 있으며 때때로 기능성을 향상시키기 위해 아미노산 시퀀스를 합성/변형하여 사용된다. 상기 EEPs는 일반적으로 30개 이하의 아미노산으로 이루어져 있는 반면에, BoNT/A 트렌스로케이션 도메인의 아미노산 수는 435개 이므로, 기존 보툴리눔 신경독소의 트렌스로케이션 도메인을 대체하기 위한 목적으로 도입할 경우 기존 보다 훨씬 작은 분자크기의 보툴리눔 신경독소 미니어처를 기대할 수 있다.When BoNT is attached to the presynaptic membrane and is absorbed into the cell, it is absorbed by forming a vesicle from the presynaptic membrane. In order for BoNTs to inhibit SNARE complex formation, escape from the vesicle lumen to the cytoplasm is required, and the part that moves BoNT LCs from the vesicle to the cytoplasm is the translocation domain of HC. When the pH in the vesicle decreases, alpha-helices in the translocation domain form a channel in the vesicle membrane, and after the LC passes through the channel and moves in the cytoplasmic direction, the disulfide bond with HC is broken It is present in an activated form in the cytoplasm. Since the recombinant botulinum neurotoxin form proposed in the present invention assumes an activity similar to that of the existing botulinum neurotoxin, a portion having a function (vesicle escape) similar to the translocation domain of the BoNT is required. Various peptides (endosomal escape peptides, EEPs) that help escape vesicles and move to the cytoplasm are mainly known from natural origins such as viruses, bacteria, animals, and plants, and are sometimes used by synthesizing/modifying amino acid sequences to improve functionality. . While the EEPs generally consist of 30 or less amino acids, the number of amino acids in the BoNT/A translocation domain is 435. Therefore, when introduced for the purpose of replacing the translocation domain of the existing botulinum neurotoxin, it is much more than before. A miniature botulinum neurotoxin with a small molecular size can be expected.
본 발명에서 상기 BoNTs LC는 서열번호 1 내지 70으로 이루어진 군으로부터 선택되는 아미노산 서열을 가질 수 있으나 이에 제한되는 것은 아니다.In the present invention, the BoNTs LC may have an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 70, but is not limited thereto.
본 발명에서 상기 EEP는 서열번호 76 내지 98로 이루어진 군으로부터 선택되는 아미노산 서열을 가질 수 있으나 이에 제한되는 것은 아니다.In the present invention, the EEP may have an amino acid sequence selected from the group consisting of SEQ ID NOs: 76 to 98, but is not limited thereto.
본 발명에서 상기 BoNT/A-A2는 서열번호 116 내지 120으로 이루어진 군으로부터 선택되는 아미노산 서열을 가질 수 있으나 이에 제한되는 것은 아니다.In the present invention, the BoNT / A-A2 may have an amino acid sequence selected from the group consisting of SEQ ID NOs: 116 to 120, but is not limited thereto.
본 발명의 다른 양태로서, 본 발명은 상기 SNARE(Soluble Nethylameimide-sensitive factor Attachment Protein Receptor; SNAP Receptor) 복합체 형성 저해용 조성물을 유효성분으로 포함하는, 신경병증 예방 또는 치료용 약학적 조성물을 제공한다.As another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating neuropathy, comprising the composition for inhibiting the formation of the SNARE (Soluble Nethylameimide-sensitive factor Attachment Protein Receptor; SNAP Receptor) complex as an active ingredient.
본 발명에서 사용되는 용어, "예방"이란 본 발명에 따른 약학적 조성물의 투여에 의해 신경병증을 억제하거나 발병을 지연시키는 모든 행위를 의미한다.As used herein, the term “prevention” refers to any action that suppresses or delays the onset of neuropathy by administration of the pharmaceutical composition according to the present invention.
본 발명에서 사용되는 용어 "치료"는, 본 발명에 따른 약학적 조성물의 투여에 의해 신경병증에 의한 증세가 호전되거나 이롭게 되도록 하는 모든 행위를 의미한다.As used herein, the term “treatment” refers to any action that improves or benefits symptoms due to neuropathy by administration of the pharmaceutical composition according to the present invention.
본 발명에 따른 약학적 조성물은 상기 SNARE 복합체 형성 저해용 조성물을 유효성분으로 포함하며, 약학적으로 허용 가능한 담체를 포함할 수 있다. 상기 약학적으로 허용 가능한 담체는 제제시에 통상적으로 이용되는 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 사이클로덱스트린, 덱스트로즈 용액, 말토덱스트린 용액, 글리세롤, 에탄올, 리포좀 등을 포함하지만 이에 한정되지 않으며, 필요에 따라 항산화제, 완충액 등 다른 통상의 첨가제를 더 포함할 수 있다. 또한, 희석제, 분산제, 계면활성제, 결합제, 윤활제 등을 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립, 또는 정제로 제제화할 수 있다. 적합한 약학적으로 허용되는 담체 및 제제화에 관해서는 레밍턴의 문헌에 개시되어 있는 방법을 이용하여 각 성분에 따라 바람직하게 제제화할 수 있다. 본 발명의 약학적 조성물은 제형에 특별한 제한은 없으나 주사제, 흡입제, 피부 외용제, 또는 경구 섭취제 등으로 제제화할 수 있다.The pharmaceutical composition according to the present invention includes the composition for inhibiting SNARE complex formation as an active ingredient, and may include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is commonly used in formulation, and includes, but is not limited to, saline, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposome, and the like. It does not, and may further include other conventional additives such as antioxidants and buffers, if necessary. In addition, diluents, dispersants, surfactants, binders, lubricants, etc. may be additionally added to form an injectable formulation such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules, or tablets. Regarding suitable pharmaceutically acceptable carriers and formulations, formulations can be preferably made according to each component using the method disclosed in Remington's literature. The pharmaceutical composition of the present invention is not particularly limited in the formulation, but may be formulated as an injection, an inhalant, an external preparation for the skin, or an oral intake.
본 발명의 약학적 조성물은 목적하는 방법에 따라 경구 투여하거나 비경구투여(예를 들어, 정맥 내, 피하, 피부, 비강, 기도에 적용)할 수 있으며, 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 시간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다.The pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intravenously, subcutaneously, dermally, nasally, applied to the respiratory tract) according to a desired method, and the dosage may vary depending on the patient's condition and body weight, disease Although it varies depending on the degree, drug form, administration route, and time of administration, it may be appropriately selected by those skilled in the art.
본 발명에 따른 조성물은 약학적으로 유효한 양으로 투여한다. 본 발명에 있어서, "약학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효용량 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명에 따른 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type, severity, and drug activity of the patient. , sensitivity to drug, administration time, administration route and excretion rate, duration of treatment, factors including concurrent drugs, and other factors well known in the medical field. The composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.
구체적으로, 일일 투여량은 상기 조성물의 양을 기준으로 0.01 내지 10 ㎎/㎏이고, 바람직하게는 0.1 내지 1 ㎎/㎏이며, 하루 1 내지 6 회 투여될 수 있다. 또한, 그 투여량은 연령, 성별, 체중, 질병의 정도, 투여경로 등에 따라서 증감될 수 있다. 따라서 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.Specifically, the daily dose is 0.01 to 10 mg/kg, preferably 0.1 to 1 mg/kg, based on the amount of the composition, and may be administered 1 to 6 times a day. In addition, the dosage may be increased or decreased according to age, sex, weight, degree of disease, administration route, and the like. Therefore, the above dosage does not limit the scope of the present invention in any way.
본 발명에서 상기 신경병증은 사시, 안검경련, 성대장애, 사경, 심근장애, 궤양, 위산역류질환, 식욕감소, 췌장질환, 튼 살, 절박성 요실금, 치열, 소아마비, 근육통, 엉덩이 기형, 다한증, 허리통증, 경부통, 만성두통 및 뇌신경장애로 이루어진 군으로부터 선택될 수 있으나 이에 제한되는 것은 아니다.In the present invention, the neuropathy is strabismus, blepharospasm, vocal cord disorder, torticollis, myocardial disorder, ulcer, acid reflux disease, decreased appetite, pancreatic disease, stretch marks, urge incontinence, dentition, polio, myalgia, hip deformity, hyperhidrosis, lower back It may be selected from the group consisting of pain, neck pain, chronic headache, and cranial nerve disorder, but is not limited thereto.
본 발명의 다른 양태로서, 상기 SNARE(Soluble Nethylameimide-sensitive factor Attachment Protein Receptor; SNAP Receptor) 복합체 형성 저해용 조성물을 유효성분으로 포함하는, 화장료 조성물을 제공한다.As another aspect of the present invention, there is provided a cosmetic composition comprising the SNARE (Soluble Nethylameimide-sensitive factor Attachment Protein Receptor; SNAP Receptor) complex formation inhibitory composition as an active ingredient.
본 발명에서 상기 화장료 조성물은 주름방지 효과가 있을 수 있다.In the present invention, the cosmetic composition may have an anti-wrinkle effect.
피부의 모공은 얼굴 피부에 전체적으로 고르게 분포하며, 모공 내에 존재하는 털과 붙어있는 근육(입모근)에 의해 여닫음이 조절되고 해당 입모근의 수축 또는 이완은 각각 교감신경과 부교감신경의 지배를 받으며, 구체적으로, 부교감신경은 입모근 주위에 존재하여 모공의 확대 작용을 유도하고, 교감신경은 입모근과의 물리적 거리가 비교적 멀고 모공의 축소 작용을 유도한다. 따라서 선택적으로 모공근처에 존재하는 부교감 신경만을 억제하면 보상 작용으로 교감신경만이 작용하게 되어 털에 붙어 있는 근육을 수축시켜 모공이 축소될 수 있다. 이와 더불어 과도한 얼굴근육 운동에 의한 주름의 생성을 억제하고, 피부를 당겨지는 효과로 인해 생성된 주름을 개선할 수 있다. 기존 상용되는 보톡스 또한 해당 기작을 통해 피부미용을 목적으로 사용되고 있으며, 주름 및 모공 개선효과가 있음이 이미 확인되었다.The pores of the skin are evenly distributed throughout the skin of the face, opening and closing is controlled by the muscles attached to the hairs in the pores (the trapezius muscle), and the contraction or relaxation of the trapezius muscle is controlled by the sympathetic nerve and the parasympathetic nerve, respectively. Therefore, the parasympathetic nerve exists around the trapezius muscle and induces the enlargement of the pores, and the sympathetic nerve has a relatively long physical distance from the trapezius and induces the contraction of the pores. Therefore, if you selectively suppress only the parasympathetic nerves that exist near the pores, only the sympathetic nerves act as a compensatory action, thereby contracting the muscles attached to the hair, thereby reducing the pores. In addition, it is possible to suppress the generation of wrinkles due to excessive facial muscle exercise and to improve the wrinkles generated due to the effect of pulling the skin. Existing commercial Botox is also used for skin beauty through the mechanism, and it has already been confirmed that it has the effect of improving wrinkles and pores.
본 발명의 상기 화장료 조성물은 상기 SNARE 복합체 형성 저해용 조성물뿐만 아니라, 화장료 조성물에 통상적으로 이용되는 성분들을 포함할 수 있으며, 예컨대 항산화제, 안정화제, 용해화제, 비타민, 안료, 및 향료와 같은 통상적인 보조제, 그리고 담체를 포함할 수 있다. The cosmetic composition of the present invention may include not only the composition for inhibiting the formation of the SNARE complex, but also components commonly used in cosmetic compositions, for example, conventional antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances. phosphorus adjuvants, and carriers.
또한, 본 발명의 조성물은 상기 SNARE 복합체 형성 저해용 조성물 이외에, 상기 SNARE 복합체 형성 저해용 조성물과 반응하여 피부보호 효과를 손상시키지 않는 한도에서 종래부터 사용되어오던 유기 자외선 차단제를 혼합하여 사용할 수도 있다. 상기 유기 자외선 차단제로는 글리세릴파바, 드로메트리졸트리실록산, 드로메트리졸, 디갈로일트리올리에이트, 디소듐페닐디벤즈이미다졸테트라설포네이트, 디에틸헥실부타미도트리아존, 디에틸아미노하이드록시벤조일헥실벤조에이트, 디이에이-메톡시신나메이트, 로우손과 디하이드록시아세톤의 혼합물, 메틸렌비스-벤조트리아졸릴테트라메칠부틸페놀, 4-메틸벤질리덴캠퍼, 멘틸안트라닐레이트, 벤조페논-3(옥시벤존), 벤조페논-4, 벤조페논-8(디옥시페벤존), 부틸메톡시디벤조일메탄, 비스에틸헥실옥시페놀메톡시페닐트리아진, 시녹세이트, 에틸디하이드록시프로필파바, 옥토크릴렌, 에틸헥실디메틸파바, 에틸헥실메톡시신나메이트, 에틸헥실살리실레이트, 에틸헥실트리아존, 이소아밀-p-메톡시신나메이트, 폴리실리콘-15(디메치코디에틸벤잘말로네이트), 테레프탈릴리덴디캠퍼설포닉애씨드 및 그 염류, 티이에이-살리실레이트 및 아미노벤조산(파바)으로 이루어진 군으로부터 선택된 1종 이상을 사용할 수 있다. In addition, the composition of the present invention, in addition to the composition for inhibiting the formation of the SNARE complex, may be used by mixing an organic sunscreen that has been conventionally used as long as it does not damage the skin protection effect by reacting with the composition for inhibiting the formation of the SNARE complex. Examples of the organic sunscreen include glyceryl pava, dromethrizole trisiloxane, dromethrazole, digalloyl trioleate, disodium phenyldibenzimidazole tetrasulfonate, diethylhexylbutamidotriazone, diethylamino Hydroxybenzoylhexylbenzoate, DEA-methoxycinnamate, Lawson and dihydroxyacetone mixture, methylenebis-benzotriazolyltetramethylbutylphenol, 4-methylbenzylidene camphor, menthylanthranilate, benzophenone -3 (oxybenzone), benzophenone-4, benzophenone-8 (deoxyphenzone), butylmethoxydibenzoylmethane, bisethylhexyloxyphenol methoxyphenyltriazine, cinoxate, ethyldihydroxypropylfaba, Octocrylene, ethylhexyldimethylfaba, ethylhexylmethoxycinnamate, ethylhexylsalicylate, ethylhexyltriazone, isoamyl-p-methoxycinnamate, polysilicon-15 (dimethicodiethylbenzalmalonate) , terephthalylidene dicamphor sulfonic acid and its salts, TEA-salicylate, and at least one selected from the group consisting of aminobenzoic acid (Fava) may be used.
본 발명의 화장료 조성물을 첨가할 수 있는 제품으로는, 예를 들어, 수렴화장수, 유연화장수, 영양화장수, 각종 크림, 에센스, 팩, 파운데이션 등과 같은 화장품류와 클렌징, 세안제, 비누, 트리트먼트, 미용액 등이 있다. 본 발명의 화장료 조성물의 구체적인 제형으로서는 스킨로션, 스킨 소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐 로션, 영양로션, 마사지크림, 영양크림, 모이스쳐 크림, 핸드크림, 에센스, 영양에센스, 팩, 비누, 샴푸, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션, 바디클렌저, 유액, 립스틱, 메이크업 베이스, 파운데이션, 프레스파우더, 루스파우더, 아이섀도 등의 제형을 포함한다.Products to which the cosmetic composition of the present invention can be added include, for example, cosmetics such as astringent lotion, softening lotion, nutritional lotion, various creams, essences, packs, foundations, and cleansing, face wash, soap, treatment, and serum. etc. Specific formulations of the cosmetic composition of the present invention include skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, essence, nourishing essence, pack, It includes formulations such as soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, lipstick, makeup base, foundation, press powder, loose powder, and eye shadow.
본 발명의 바람직한 구현예에 따르면, 본 발명의 SNARE 복합체 형성 저해용 조성물의 함량은 전체 조성물 총 중량 대비 0.001 내지 10 중량% 포함하며, 바람직하게는 0.005 내지 5 중량%, 더욱 바람직하게는 0.01 내지 3 중량% 포함할 수 있다. 상기 SNARE 복합체 형성 저해용 조성물의 함량이 0.001 중량% 미만일 경우, 피부주름 개선 효과를 기대하기 어려울 수 있으며, 10 중량%를 초과하는 경우, 조성물을 적절한 제형으로 제조하거나 장기 안정성의 확보에도 어려움이 있다.According to a preferred embodiment of the present invention, the content of the composition for inhibiting SNARE complex formation of the present invention comprises 0.001 to 10% by weight, preferably 0.005 to 5% by weight, more preferably 0.01 to 3% by weight relative to the total weight of the total composition. % by weight. If the content of the composition for inhibiting SNARE complex formation is less than 0.001% by weight, it may be difficult to expect an effect of improving skin wrinkles, and if it exceeds 10% by weight, it is difficult to prepare the composition in an appropriate formulation or to secure long-term stability. .
본 발명의 또 다른 양태로서, 뉴런 표시용 재조합 형광단백질을 포함하는 조성물을 제공한다.As another aspect of the present invention, there is provided a composition comprising a recombinant fluorescent protein for neuronal display.
본 발명에서 상기 뉴런 표시용 재조합 형광단백질은 녹색, 노란색, 청록색, 빨간색, 파란색형광단백질일 수 있으나, 상기 색으로 정의되지 않는 특정한 파장대의 형광을 발광하는 형광단백질일 수 있으며, 또한 단백질이 아닌 저분자 발광화합물일 수 있다.In the present invention, the recombinant fluorescent protein for neuron display may be a green, yellow, cyan, red, or blue fluorescent protein, but may be a fluorescent protein that emits fluorescence in a specific wavelength band not defined by the color, and is not a protein but a small molecule It may be a luminescent compound.
본 발명에서 상기 뉴런 표시용 재조합 형광단백질은 실험적 또는 의학적 용도로 사용될 수 있으나, 이에 국한되는 것은 아니다.In the present invention, the recombinant fluorescent protein for neuron display may be used for experimental or medical purposes, but is not limited thereto.
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예를 제시한다. 그러나 하기의 실시예는 본 발명을 보다 쉽게 이해하기 위하여 제공되는 것일 뿐, 하기 실시예에 의해 본 발명의 내용이 한정되는 것은 아니다.Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.
[[ 실시예Example ]]
실시예Example 1. 재조합 보툴리눔 1. Recombinant Botulinum 신경독소를neurotoxin 생산하기 위한 플라스미드의 제조 Preparation of plasmids for production
BoNTs LC 단백질을 생산하기 위한 플라스미드(1)의 제조는 pET28b 벡터를 이용하여 T4 DNA 폴리머레이즈(polymerase)를 이용한 방법으로 클로닝 하였다. 상기 클로닝 과정 중 사용된 벡터맵은 도 3과 같으며, 인설트(insert) DNA 시퀀스는 하기 표 1의 아미노산 시퀀스를 바탕으로 https://sg.idtdna.com/CodonOpt 내 Codon optimization tool의 코돈최적화(E. coli 기준) 과정을 통해 하기 표 2에 기재된 DNA 서열을 이용하였다.The plasmid (1) for producing BoNTs LC protein was cloned using a pET28b vector and T4 DNA polymerase. The vector map used during the cloning process is shown in FIG. 3, and the insert DNA sequence is based on the amino acid sequence of Table 1 below. Codon optimization of the codon optimization tool in https://sg.idtdna.com/CodonOpt ( Based on E. coli ) The DNA sequences shown in Table 2 below were used through the process.
(50 kDa)437 amino acids
(50 kDa)
(52 kDa)451 amino acids
(52 kDa)
(54 kDa)474 amino acids
(54 kDa)
(54 kDa)474 amino acids
(54 kDa)
보다 자세하게는, 최초 PCR(polymerase chain reaction)은 pET28b와 BoNTs LC 인설트의 양 말단 15 bp에 상호보완적으로 디자인된 프라이머들을 이용해 수행되었다. 그리고 PCR 완료 후에는 셀프-라이게이션(self-ligation) 방지를 위해 37 ℃에서 1 시간 동안 Dpn1 용액을 처리하여, DNA 외의 PCR 산물을 제거하였다. PCR을 통하여 증폭된 pET28b와 BoNTs LC 인설트는 라이게이션(ligation)하기 위해 T4 DNA 폴리머레이즈를 첨가하고 상온에서 2 분 30 초, 얼음에서 10 분 동안 반응시켜 pET28b와 BoNTs LC 인설트 간의 안정적인 수소결합을 유도하였다. 상기 클로닝 과정을 통해 얻어진 DNA 용액 4 μL를 competent E. coli TOP10이 포함되어 있는 용액 100 μL에 첨가하고 얼음에서 30 분 배양 후 42 ℃에서 45 초간 열처리하여 형질전환하였다. 그리고 상기 형질전환된 E. coli 용액 내에 LB(Luria-Bertani) 액체 배지 900 μL를 넣어 배양(37 ℃, 1 시간)한 후 원심분리(13,000 rpm, 10 분)를 통해 세포를 수집하였다. 수집된 E. coli 용액 중 0.1 mL을 카나마이신이 첨가된 LB 고체배지에 도말/배양(37 ℃)한 후 형성된 콜로니 중 하나를 0.1% 카나마이신이 첨가된 10 mL LB 액체 배지에 접종하고 18 시간(37 ℃) 동안 배양하였다. 그리고 배양액을 초음파 처리 및 정제하여 결과물로서의 플라스미드를 얻었다. 상기의 플라스미드의 시퀀스는 업체(바이오닉스, 한국)를 통해 확인하였다. 상기 과정에서 사용된 프라이머의 서열, 자세한 조성과 반응 조건은 이하와 같다.In more detail, the initial PCR (polymerase chain reaction) was performed using primers designed to complement each other 15 bp at both ends of pET28b and BoNTs LC Insult. And after PCR was completed, the Dpn1 solution was treated at 37° C. for 1 hour to prevent self-ligation, and PCR products other than DNA were removed. For pET28b and BoNTs LC strains amplified through PCR, T4 DNA polymerase was added for ligation and reacted for 2 minutes and 30 seconds at room temperature and 10 minutes on ice to ensure stable hydrogen bonding between pET28b and BoNTs LC strains. induced. 4 μL of the DNA solution obtained through the cloning process was added to 100 μL of a solution containing competent E. coli TOP10, incubated on ice for 30 minutes, and then heat treated at 42° C. for 45 seconds for transformation. Then, 900 μL of LB (Luria-Bertani) liquid medium was added to the transformed E. coli solution and cultured (37° C., 1 hour), and then the cells were collected by centrifugation (13,000 rpm, 10 minutes). 0.1 mL of the collected E. coli solution was smeared/cultured on LB solid medium supplemented with kanamycin (37 ° C), and one of the colonies formed was inoculated into 10 mL LB liquid medium supplemented with 0.1% kanamycin for 18 hours (37 ℃) for incubation. Then, the culture medium was subjected to sonication and purification to obtain a plasmid as a result. The sequence of the above plasmid was confirmed through a company (Bionics, Korea). The sequence, detailed composition and reaction conditions of the primers used in the above process are as follows.
- 벡터(pET28b, 5340 bp)- vector (pET28b, 5340 bp)
① 전방 프라이머(Forward primer)① Forward primer
5‘-CACCACCACCACCACCACTGAGATC-3’(서열번호 133)5'-CACCACCACCACCACCACTGAGATC-3' (SEQ ID NO: 133)
② 후방 프라이머(Backward primer)② Backward primer
5‘-CTCGCTACCGCCTCCACCACTG-3’ (서열번호 134)5'-CTCGCTACCGCCTCCACCACTG-3' (SEQ ID NO: 134)
- 재조합 보툴리눔 신경독소 인설트(BoNTs LC)- Recombinant Botulinum Neurotoxin Insult (BoNTs LC)
① 전방 프라이머① Front primer
5‘-GTGGTGGTGGTGGTGGAAGGGAATGATGCCACGGACACAAAGTAACTT-3’ (서열번호 135)5'-GTGGTGGTGGTGGTGGAAGGGAATGATGCCACGGACACAAAGTAACTT-3' (SEQ ID NO: 135)
② 후방 프라이머② Rear primer
5‘-GGAGGCGGTAGCGAGATGCCCTTCGTCAACAAACAGTTCAATTATAAGGATCC-3’ (서열번호 136)5'-GGAGGCGGTAGCGAGATGCCCTTCGTCAACAAACAGTTCAATTATAAGGATCC-3' (SEQ ID NO: 136)
- PCR 조성물: 전방 프라이머 1 μL, 후방 프라이머 1 μL, 벡터 1 μL, dNTP 4 μL, 10X 리액션 버퍼 5 μL, Pfu DNA 폴리머레이즈 0.5 μL, 증류수 37.5 μL-PCR composition: 1 μL of forward primer, 1 μL of back primer, 1 μL of vector, 4 μL of dNTP, 5 μL of 10X reaction buffer, 0.5 μL of Pfu DNA polymerase, 37.5 μL of distilled water
- 벡터의 PCR 조건- PCR conditions of the vector
① 최초 변성(denaturation, 95℃, 3분)① Initial denaturation (95℃, 3 minutes)
② 30회 반복: 변성(95℃, 3초) → 결합(annealing, 61℃, 3초) → 연장(extension, 72℃, 6분)②
③ 마지막 연장(extension, 72℃, 3분)③ Last extension (extension, 72℃, 3 minutes)
- 인설트의 PCR 조건- PCR conditions of Insult
① 최초 변성(95℃, 3분)① Initial denaturation (95℃, 3 minutes)
② 30회 반복: 변성(95℃, 3초) → 결합(61℃, 3초) → 연장(72℃, 1분)②
③ 마지막 연장(72℃, 3분)③ Last extension (72℃, 3 minutes)
- Dpn1 처리조건(37℃, 1시간): 5 μL(10X reaction buffer4) + 44 μL(벡터 또는 인설트의 PCR 산물) + 1 μL(Dpn1 용액)- Dpn1 treatment conditions (37℃, 1 hour): 5 μL (10X reaction buffer4) + 44 μL (PCR product of vector or insult) + 1 μL (Dpn1 solution)
- T4 DNA 폴리머레이즈 처리: 1 μL(벡터 용액) + 7 μL(인설트 용액) + 1 μL(10X reaction buffer2.1) + 1 μL(T4 DNA polymerase 용액)- T4 DNA polymerase treatment: 1 μL (vector solution) + 7 μL (insult solution) + 1 μL (10X reaction buffer2.1) + 1 μL (T4 DNA polymerase solution)
BoNTs LC 이외의 도 2의 (2) 내지 (6)에서 나타난 재조합 보툴리눔 신경독소의 생산에서, Aurein 1.2(서열번호 92), His10(서열번호 76), A2(서열번호 156)에 해당하는 각각의 서열은 비교적 짧기 때문에 T4 DNA 폴리머레이즈를 이용한 방법이 아닌 부위 특이적 변이(site-directed mutagenesis) 방법으로 특정 DNA에 상기의 짧은 특정서열이 삽입되도록 하였으며 특히 전방 프라이머(forward primer)는 앞 말단에 희망서열이 삽입되도록 디자인하였다. BoNTs LC 생산용 플라스미드(1)에 기초하여, 상기 부위 특이적 변이 방법으로 BoNTs LC의 C 말단에 A2가 삽입되도록 재조합 보툴리눔 신경독소(BoNTs LC-A2) 생산용 플라스미드(2)를 디자인하였다. 상기 플라스미드(2)를 이용하여 BoNTs LC와 A2 사이에 Aurein 1.2가 삽입되도록 플라스미드(3)(BoNTs LC-Aurein 1.2-A2)와, BoNTs LC와 A2 사이에 His10이 삽입되도록 플라스미드(4)(BoNTs LC-His10-A2)를 디자인하였다. 그리고 상기 플라스미드(4)를 이용하여 BoNTs LC의 N 말단에 Aurein 1.2가 삽입되도록 플라스미드(5)(Aurein 1.2-BoNTs LC-His10-A2)를 디자인하였다. 마지막으로, 플라스미드(2)를 이용하여 BoNTs LC의 N 말단에 His10과 Aurein1.2가 차례로 삽입될 수 있도록 플라스미드(6)(Aurein 1.2-His10-LC-A2)을 디자인하였다. 상기 과정에서 사용된 각각의 프라이머 염기서열은 다음과 같다. In the production of recombinant botulinum neurotoxin shown in (2) to (6) of FIG. 2 other than BoNTs LC, each corresponding to Aurein 1.2 (SEQ ID NO: 92), His10 (SEQ ID NO: 76), A2 (SEQ ID NO: 156) Since the sequence is relatively short, the short specific sequence was inserted into a specific DNA by a site-directed mutagenesis method rather than a method using T4 DNA polymerase. In particular, a forward primer is desired at the front end The sequence was designed to be inserted. Based on the plasmid for BoNTs LC production (1), a recombinant botulinum neurotoxin (BoNTs LC-A2) production plasmid (2) was designed such that A2 was inserted into the C terminus of BoNTs LC by the site-specific mutation method. Plasmid (3) (BoNTs LC-Aurein 1.2-A2) to insert Aurein 1.2 between BoNTs LC and A2 using the above plasmid (2), and plasmid (4) (BoNTs) to insert His10 between BoNTs LC and A2 LC-His10-A2) was designed. Then, using the plasmid (4), a plasmid (5) (Aurein 1.2-BoNTs LC-His10-A2) was designed so that Aurein 1.2 was inserted into the N-terminus of the BoNTs LC. Finally, plasmid (6) (Aurein 1.2-His10-LC-A2) was designed so that His10 and Aurein1.2 could be sequentially inserted into the N-terminus of BoNTs LC using plasmid (2). Each primer base sequence used in the above process is as follows.
- 재조합 보툴리눔 신경독소(BoNTs LC-A2)- Recombinant botulinum neurotoxin (BoNTs LC-A2)
① 전방 프라이머① Front primer
5‘-TGTGCTGTTCAAATAGATGTTTGTGGTCACAACGCTCCCGCGAGAGGGTCTGTGGTCACCACAAATATATACCTGA-3’ (서열번호 137)5'-TGTGCTGTTCAAATAGATGTTTGTGGTCACAACGCTCCCGCGAGAGGGTCTGTGGTCACCACAAATATATACCTGA-3' (SEQ ID NO: 137)
② 후방 프라이머② Rear primer
5‘-CACCACCACCACCACCACTGAGATC-3’ (서열번호 138)5'-CACCACCACCACCACCACTGAGATC-3' (SEQ ID NO: 138)
- 재조합 보툴리눔 신경독소(BoNTs LC-Aurein 1.2-A2)- Recombinant botulinum neurotoxin (BoNTs LC-Aurein 1.2-A2)
① 전방 프라이머① Front primer
5‘-AAATGATTCTGCGATCTTTATTATGATGTCAAACAGTCCAGAGGGTCTGTGGTCACCACAAATATATACCTG-3’ (서열번호 139)5'-AAATGATTCTGCGATCTTTATTATGATGTCAAACAGTCCAGAGGGTCTGTGGTCACCACAAATATATAACCTG-3' (SEQ ID NO: 139)
② 후방 프라이머② Rear primer
5‘-CGCGGGAGCGTTGTGACCACAAACAT-3’ (서열번호 140)5'-CGCGGGAGCGTTGTGACCACAAACAT-3' (SEQ ID NO: 140)
- 재조합 보툴리눔 신경독소(BoNTs LC-His10-A2)- Recombinant botulinum neurotoxin (BoNTs LC-His10-A2)
① 전방 프라이머① Front primer
5‘-GTGATGGTGATGGTGATGGTGATGGTGGTGAGAGGGTCTGTGGTCACCACAAATATATACCTG-3’ (서열번호 141)5'-GTGATGGTGATGGTGATGGTGATGGTGGTGAGAGGGTCTGTGGTCACCACAAATATATAACCTG-3' (SEQ ID NO: 141)
② 후방 프라이머② Rear primer
5‘-CGCGGGAGCGTTGTGACCACAAACAT-3’ (서열번호 142)5'-CGCGGGAGCGTTGTGACCACAAACAT-3' (SEQ ID NO: 142)
- 재조합 보툴리눔 신경독소(Aurein 1.2-BoNTs LC-His10-A2)- Recombinant botulinum neurotoxin (Aurein 1.2-BoNTs LC-His10-A2)
① 전방 프라이머① Front primer
5‘-AAATGATTCTGCGATCTTTATTATGATGTCAAACAGCTCGCTACCGCCTCCACCACTG-3’ (서열번호 143)5'-AAATGATTCTGCGATCTTTATTATGATGTCAAACAGCTCGCTACCGCCTCCACCACTG-3' (SEQ ID NO: 143)
② 후방 프라이머② Rear primer
5‘-ATGCCCTTCGTCAACAAACAGTTCAATTATAAGGATCC-3’ (서열번호 144)5'-ATGCCCTTCGTCAACAAACAGTTCAATTATAAGGATCC-3' (SEQ ID NO: 144)
- 재조합 보툴리눔 신경독소(Aurein 1.2-His10-BoNTs LC-A2) 1st 단계- Recombinant botulinum neurotoxin (Aurein 1.2-His10-BoNTs LC-A2) 1st step
① 전방 프라이머① Front primer
5‘-GTGATGGTGATGGTGATGGTGATGGTGGTGCTCGCTACCGCCTCCACCACTG-3’ (서열번호 145)5'-GTGATGGTGATGGTGATGGTGATGGTGGTGCTCGCTACCGCCTCCACCACTG-3' (SEQ ID NO: 145)
② 후방 프라이머② Rear primer
5‘-ATGCCCTTCGTCAACAAACAGTTCAATTATAAGGATCC-3’ (서열번호 146)5'-ATGCCCTTCGTCAACAAACAGTTCAATTATAAGGATCC-3' (SEQ ID NO: 146)
- 재조합 보툴리눔 신경독소(Aurein 1.2-His10-BoNTs LC-A2) 2nd 단계- Recombinant botulinum neurotoxin (Aurein 1.2-His10-BoNTs LC-A2) 2nd step
① 전방 프라이머① Front primer
5‘-AAATGATTCTGCGATCTTTATTATGATGTCAAACAGTCCCTCGCTACCGCCTCCACCACTG-3’ (서열번호 147)5'-AAATGATTCTGCGATCTTTATTATGATGTCAAACAGTCCCTCGCTACCGCCTCCACCACTG-3' (SEQ ID NO: 147)
② 후방 프라이머② Rear primer
5‘-CACCACCATCACCATCACCATCACCATC-3’ (서열번호 148)5'-CACCACCATCACCATCACCATCACCATC-3' (SEQ ID NO: 148)
※ 모든 프라이머들의 Tm값은 65 ℃※ Tm value of all primers is 65 ℃
이후의 상기 플라스미드 결과물(2) 내지 (6)을 제조하기 위한 클로닝 방법은 상기 에 기재된 BoNTs LC 생산용 플라스미드(1)의 제조방법과 동일한 방법으로 수행하였다. 자세한 조성과 반응 조건은 다음과 같다.The cloning method for the subsequent preparation of the plasmid products (2) to (6) was performed in the same manner as the preparation method of the plasmid (1) for producing BoNTs LC described above. Detailed composition and reaction conditions are as follows.
- PCR 조건- PCR conditions
① 최초 변성(95℃, 3분)① Initial denaturation (95℃, 3 minutes)
② 30회 반복: 변성(95℃, 3초) → 결합(61℃, 3초) → 연장(72℃, 6분)②
③ 마지막 연장(72℃, 3분)③ Last extension (72℃, 3 minutes)
- 라이게이션(25℃, 2시간): 5 μL(X10 ligase 버퍼) + 5 μL(10X kinase 버퍼) + 10 μL(PCR 산물) + 0.5 μL(ligase 용액) + 1 μL(kinase 용액)- Ligation (25℃, 2 hours): 5 μL (X10 ligase buffer) + 5 μL (10X kinase buffer) + 10 μL (PCR product) + 0.5 μL (ligase solution) + 1 μL (kinase solution)
※ PCR 조성 및 Dpn1 처리조건은 상기 플라스미드(1) 제작방법과 상동※ PCR composition and Dpn1 treatment conditions are the same as the above plasmid (1) production method
※ Dpn1, 10X reaction buffer, 10X reaction buffer4, 10X reaction buffer2.1, Pfu DNA polymerase T4 DNA polymerase, dNTP(2.5mM)의 구입처: ELPIS BIOTECH, 한국※ Where to purchase Dpn1, 10X reaction buffer, 10X reaction buffer4, 10X reaction buffer2.1, Pfu DNA polymerase T4 DNA polymerase, dNTP (2.5mM): ELPIS BIOTECH, Korea
※ PCR 산물의 정제법 및 플라스미드의 제조법: Dokdo-prep Gel extraction kit spintype 200 (ELPIS)에 내장된 프로토콜에 따름※ PCR product purification and plasmid preparation: Follow the protocol built in Dokdo-prep Gel extraction kit spintype 200 (ELPIS)
실시예Example 2. E. 2. E. coli를coli 이용한 보툴리눔 botulinum using 신경독소neurotoxin 미니어처 생산 miniature production
실시예 1에서 제조된 플라스미드(1) 내지 (6)을 이용하여 재조합 보툴리눔 신경독소를 생산하는 방법은 이하와 같다. 수용성 세포(competent cell)로 변환된 E. coli BL21(DE3) 균주에 전기에 의한 천공을 통하거나 열 충격을 가함으로써 상기 완성된 플라스미드를 균주 내부로 이동시킴으로써 형질 도입시켰다. 그리고 형질 도입된 E. coli를 암피실린이 첨가된 LB 고체 선택배지에 도말한 후 37 ℃에서 하루 동안 배양하였으며, 선택배지에서 자란 1 개의 콜로니를 암피실린이 첨가된 액체 LB 배지 10 mL에 접종한 후 12 시간 동안 37 ℃에서 전 배양하였다. A method for producing a recombinant botulinum neurotoxin using the plasmids (1) to (6) prepared in Example 1 is as follows. E. coli BL21 (DE3) strain transformed into competent cells was transduced by transferring the completed plasmid into the strain by electroporation or heat shock. Then, the transduced E. coli was smeared on LB solid selective medium supplemented with ampicillin and cultured for one day at 37 ° C. One colony grown in the selective medium was inoculated into 10 mL of ampicillin-added liquid LB medium 12 Pre-incubation at 37 °C for hours.
그 뒤, 전 배양한 재조합 균주를 다시 암피실린이 첨가된 600 mL의 LB 배지에 1 %의 농도로 접종하고, OD600nm 값이 0.5가 될 때까지 37 ℃에서 배양하였다. 그리고, 상기 배양액을 8,000 rpm, 4 ℃에서 10 분 동안 원심분리하여 배지를 제거하고 균주들을 수확하였으며, 수확한 균주들은 10 mL의 PBS(phosphate buffered saline; 137 mM NaCl, 2.7 mM KCl, 2.55 mM Na2HPO4, 1.47 mM KH2PO4)에 재부유시킨 후 1 초 간격으로 1 분 45 초 간 초음파 처리(sonication)하여 세포를 파쇄하였다. 그리고 다시 4 ℃, 13,000 rpm에서 10 분간 원심분리하여 침전물을 제거하고 상층액을 획득하였다. 그리고 획득된 상층액 내에 부유하는, 발현된 재조합 보툴리눔 신경독소를 분리하기 위해, Ni-NTA 비드(bead)가 담긴 4 ℃ 컬럼 내에서 2 시간동안 장동(nutation)운동 시킴으로써 재조합 보툴리눔 신경독소의 His-tag을 통해 비드에 부착되도록 하였다. 상기 장동운동 후, PBS 10-15 mL로 세척하여 재조합 보툴리눔 신경독소를 제외한 불순물을 제거하였다. 그 다음, 상기 세척된 컬럼에 150 mM의 이미다졸(imidazole) 용액을 처리하여 단백질을 비드에서 떨어뜨림으로써 순수한 재조합 보툴리눔 신경독소를 획득하였다.Thereafter, the pre-cultured recombinant strain was again inoculated in 600 mL of LB medium supplemented with ampicillin at a concentration of 1%, and cultured at 37° C. until the OD600nm value became 0.5. Then, the culture medium was centrifuged at 8,000 rpm, 4° C. for 10 minutes to remove the medium and harvested strains. The harvested strains were 10 mL of phosphate buffered saline (PBS; 137 mM NaCl, 2.7 mM KCl, 2.55 mM Na). After resuspending in 2 HPO 4 , 1.47 mM KH 2 PO 4 ), the cells were disrupted by sonication at 1 second intervals for 1 minute and 45 seconds. Then, the precipitate was removed by centrifugation at 4 °C and 13,000 rpm for 10 minutes to obtain a supernatant. And in order to separate the expressed recombinant botulinum neurotoxin floating in the obtained supernatant, the His- of recombinant botulinum neurotoxin was performed by nutation in a column containing Ni-NTA beads at 4 ° C for 2 hours. It was attached to the beads through a tag. After the bowel movement, it was washed with 10-15 mL of PBS to remove impurities except for recombinant botulinum neurotoxin. Then, the washed column was treated with a 150 mM imidazole solution to drop the protein from the beads to obtain a pure recombinant botulinum neurotoxin.
실시예Example 3. 분화된 PC12 세포에서의 노르에피네프린 방출 분석 3. Analysis of Norepinephrine Release in Differentiated PC12 Cells
분화된 PC12 세포에서의 노르에피네프린의 방출량 분석은 세포수준에서의 신경전달저해효능을 결정하는데 사용되었다. 한국세포주은행으로부터 분양받은 랫트의 부신(rat adrenal gland) 유래의 PC12 세포(21721)를 배양접시의 70-80 %에 이를 때까지 DMEM(100 μg/mL streptomycin, 100 U/mL penicillin, 2 mM L-glutamine, and 10% heat-inactivated FBS)에서 배양하였다. 이후 신경생장인자(NGF; 7S, 50 ng/mL, Invitrogen)를 처리하고 5일 동안 추가 배양하여 분화시킨 후 분화된 PC12 세포에 high-K+ 용액(115 mM NaCl, 50 mM KCl, 1.2 mM KH2PO4, 2.5 mM CaCl2, 1.2 mM MgSO4, 11 mM glucose, and 15 mM HEPES-Tris, pH 7.4)을 처리하고 37 ℃, 5 % CO2 인큐베이터에서 15 분간 다시 배양하였다. 이후 low-K+ 용액(140 mM NaCl, 4.7 mM KCl, 1.2 mM KH2PO4, 2.5 mM CaCl2, 1.2 mM MgSO4, 11 mM glucose, and 15 mM HEPES-Tris, pH 7.4)으로 세척 후, 도 2의 (1) 내지 (6)에 나타낸 재조합 보툴리눔 신경독소 50 pM이 함유된 low-K+ 용액을 처리하였다. 이후 37 ℃, 5 % CO2 인큐베이터에서 24 시간 재배양한 후, low-K+ 용액으로 1분 동안 세척하였으며 그 뒤, high-K+ 용액을 처리하여 신경전달물질이 분비되도록 유도하였다. 분비된 신경전달물질을 함유한 완충액에서의 노르에피네프린의 농도는 도 4에 나타난 경쟁적 ELISA 기법을 사용하는 노르에피네프린(norepinephrine) ELISA 키트(KA1891, Abnova Co.)를 이용하여 정량하였다. 정량의 과정은 분화된 PC12 세포로부터 분비된 신경전달물질을 함유한 high-K+ 용액 300 μL를 extraction 96-well 플레이트로 옮긴 후부터 제조업체에서 제공된 프로토콜과 동일하게 따라 진행되었으며 최종적으로, 반응 기질을 처리하여 450 nm에서 흡광도를 측정함으로써 배출된 노르에피네프린의 양을 측정하였다.Analysis of the release amount of norepinephrine in differentiated PC12 cells was used to determine the neurotransmission inhibitory effect at the cellular level. Rat adrenal gland-derived PC12 cells (21721) purchased from the Korea Cell Line Bank were treated with DMEM (100 μg/mL streptomycin, 100 U/mL penicillin, 2 mM L) until 70-80% of the culture dish was reached. -glutamine, and 10% heat-inactivated FBS). After treatment with neurogrowth factor (NGF; 7S, 50 ng/mL, Invitrogen) and further culturing for 5 days for differentiation, the differentiated PC12 cells were treated with a high-K+ solution (115 mM NaCl, 50 mM KCl, 1.2 mM KH 2 ). PO 4 , 2.5 mM CaCl 2 , 1.2 mM MgSO 4 , 11 mM glucose, and 15 mM HEPES-Tris, pH 7.4) were treated and incubated again at 37° C., 5
그 결과, 도 5에 나타난 바와 같이 아무것도 처리하지 않은 군에 비하여 보툴리눔 신경독소 미니어처(1)은 1±3%, (2)는 16±2%, (3)은 32±10%, (4)는 31±6%, (5)는 50±6%, (6)은 61±5%의 노르에피네프린 방출을 억제하는 것으로 확인하였다. 상기 결과들을 통하여, 신경-활성 물질(neuro-active material), 즉, 보톨리눔 신경독소의 경쇄(light chain) 등과 같은 신경 내에서 작동하는 활성 물질에 EEP 및 BoNT/A-A2를 결합시킴으로써 뉴런을 특이적으로 인식하고 뉴런 내의 세포질로 탈출하여 활성(노르에피네프린 방출 저해)을 나타내는 것을 확인할 수 있었다.As a result, as shown in FIG. 5 , compared to the untreated group, the botulinum neurotoxin miniature (1) was 1±3%, (2) was 16±2%, (3) was 32±10%, (4) It was confirmed that 31±6%, (5) 50±6%, and (6) inhibited norepinephrine release by 61±5%. Through the above results, neuron by binding EEP and BoNT/A-A2 to neuro-active material, i.e., an active material acting in the nerve, such as the light chain of botulinum neurotoxin, etc. It was confirmed that it was specifically recognized and escaped into the cytoplasm within the neuron to exhibit activity (inhibition of norepinephrine release).
실시예Example 4. 4. 디지트digit 어브덕션Abduction 스코어(Digit abduction score; DAS) Digit abduction score (DAS) 어세이assay
DAS 분석은 국소 근육약화효능을 결정하는데 사용되었다. 17-21 g의 Swiss Webster 마우스를 상기 분석법에 사용했다. 실험 전 마우스는 음식과 물을 자유롭게 섭취할 수 있도록 그룹 당 5 마리씩 분류하였다. 그리고 각 마우스의 오른 비복근 위쪽 부분의 근육주사를 통해 재조합 보툴리눔 신경독소(실시예 1에서 제조된 재조합 보툴리눔 신경독소, 도 2의 (1) 내지 (6) 참조)가 투여되었다. 주사는 멸균된 250 μL 주사기에 부착된 30 게이지 바늘을 사용하여 5 μL를 각각 투여하였으며, 투여된 샘플의 용량은 10 U kg-1 체중이었으며, 각 실험은 실험군당 5 마리의 마우스를 사용하였다. 샘플을 주사하고 이틀이 지난 후 샘플이 주사된 마우스 오른 뒷다리의 발가락이 펴지지 않는 것과 다리전체가 마비된 정도를 그렇지 않은 왼 뒷다리와 비교하여 5 점 척도로 점수가 매겨졌다(0 = 정상; 1 = 펴진 발, 두 발가락이 맞닿아 있고 나머지 발가락은 펴짐; 2 = 펴진 발, 모든 발가락 끝 사이에 약간의 공간만이 존재하거나 세 발가락이 맞닿음; 3 = 발이 펴질 때 모든 발가락이 맞닿아 있거나 발이 굽을 때 네 발가락이 모임; 4 = 굽은 발, 모든 발가락이 맞닿음).DAS analysis was used to determine local muscle-weakening potency. 17-21 g Swiss Webster mice were used in this assay. Before the experiment, the mice were divided into 5 mice per group so that they could freely consume food and water. In addition, recombinant botulinum neurotoxin (recombinant botulinum neurotoxin prepared in Example 1, refer to (1) to (6) of FIG. 2 ) was administered through intramuscular injection of the upper part of the right gastrocnemius muscle of each mouse. For injection, 5 μL was administered using a 30 gauge needle attached to a sterile 250 μL syringe, and the dose of the administered sample was 10 U kg −1 body weight, and each experiment used 5 mice per experimental group. Two days after the injection of the sample, the inability to straighten the toe and the degree of paralysis of the right hind limb of the sample-injected mouse were scored on a 5-point scale compared to that of the uninjected left hind limb (0 = normal; 1 =). Feet spread out, both toes touching and remaining toes extended; 2 = Feet spread out, there is only little space between the tips of all toes or three toes are touching; 3 = All toes touching or the foot is bent when the foot is extended when all four toes meet; 4 = foot bent, all toes touching).
그 결과, 도 6에 나타난 바와 같이 DAS 어세이 상에서 보툴리눔 재조합 신경독소(1)은 0.2±0.4, (2)는 0.6±0.5, (3)은 1.8±0.8, (4)는 1.6±0.5, (5)는 2.8±0.4, (6)은 3.2±0.4로 점수 값이 매겨졌다. 상기 결과는 본 발명의 조성물이 단순히 근육을 마비시킬 수 있다는 것만을 의미하는 것이 아니라, 입모근의 움직임을 마비시킴으로써 피부주름과 모공 개선효과를 야기하는 보톡스와 동일한 기작으로 피부주름과 모공을 개선시킬 수 있다는 것을 확인할 수 있었다. 또한, 보톨리눔 신경독소 경쇄(light chain) 이외의 다양한 신경-활성 물질에 EEP 및 BoNT/A-A2를 결합시킨다면 뉴런 특이적으로 활성을 나타내는 약물로도 사용이 가능하다는 것을 확인할 수 있었다.As a result, as shown in FIG. 6 , in the DAS assay, botulinum recombinant neurotoxin (1) was 0.2±0.4, (2) was 0.6±0.5, (3) was 1.8±0.8, (4) was 1.6±0.5, ( 5) was scored as 2.8±0.4 and (6) as 3.2±0.4. The above result does not mean that the composition of the present invention can simply paralyze the muscles, but it can improve skin wrinkles and pores with the same mechanism as Botox, which causes skin wrinkles and pores improvement effect by paralyzing the movement of the pilaris muscle. could confirm that there was In addition, it was confirmed that if EEP and BoNT/A-A2 were bound to various neuro-active substances other than the botulinum neurotoxin light chain, it could be used as a neuron-specifically active drug.
실시예Example 5. 뉴런 표시용 재조합 5. Recombination for Neuron Marking 녹색형광단백질을green fluorescent protein 생산하기 위한 플라스미드의 제조 Preparation of plasmids for production
Botulinum toxin A(BoNT/A)의 receptor binding domain(RBD)의 일부 서열인 A2 펩타이드는 신경세포에 존재하는 수용체(synaptic vesicle glycoprotein 2C, SV2C)를 인식해 결합하는데 중요하게 작용하는 부분이라고 보고 되어있다(도 7A). pET28b 플라스미드 내부의 녹색형광단백질(EGFP)의 C-말단에 있는 히스티딘 테그 뒤에 링커(Gly-Gly-Gly-Gly-Ser)와 함께 펩타이드와 도메인을 클로닝하여 녹색형광단백질과 융합된 유전자를 제작하였다(도 7B). 상기 펩타이드와 결합한다고 알려진 수용체(SV2C) 또한 결합하는데 주요하게 작용하는 일부분 만을 pGex4T 플라스미드에 클로닝하여 제작하였다(도 7C). 이때 수용체와 녹색형광단백질과 융합된 유전자는 정제를 위한 서로 다른 태그를 가지도록 제작하였다. RBD을 발현하는 플라스미드의 제조는 pET28b-EGFP를 기초로 하여, SV2C를 발현하는 플라스미드의 제조는 pGex4T-1을 기초로 하여 T4 DNA polymerase를 이용한 방법으로 클로닝 하였다. 상기 클로닝 과정 중 사용된 백터맵은 도 8과 같으며, insert의 DNA 시퀀스는 하기 표 3의 아미노산 시퀀스를 바탕으로 코돈 최적화 과정을 통해 하기 표 4와 같이 얻어졌다.It has been reported that the A2 peptide, which is a part of the receptor binding domain (RBD) of botulinum toxin A (BoNT/A), plays an important role in recognizing and binding to receptors (synaptic vesicle glycoprotein 2C, SV2C) present in neurons. (Fig. 7A). A gene fused with green fluorescent protein was constructed by cloning the peptide and domain together with the linker (Gly-Gly-Gly-Gly-Ser) behind the histidine tag at the C-terminus of the green fluorescent protein (EGFP) inside the pET28b plasmid ( 7B). The receptor (SV2C), known to bind to the peptide, was also cloned into pGex4T plasmid and only a portion that plays a major role in binding (FIG. 7C). At this time, the gene fused with the receptor and green fluorescent protein was prepared to have different tags for purification. The RBD-expressing plasmid was cloned based on pET28b-EGFP, and the SV2C-expressing plasmid was cloned based on pGex4T-1 using T4 DNA polymerase. The vector map used during the cloning process is shown in FIG. 8, and the DNA sequence of the insert was obtained as shown in Table 4 below through the codon optimization process based on the amino acid sequence shown in Table 3 below.
보다 자세하게는, 최초의 PCR은 pET28b-EGFP와 RBD insert의 양 말단 15bp에 상호보완(complementary)적으로 디자인된 프라이머들을 이용해 수행되었다. 그리고 PCR 완료 후 self-ligation 방지를 위해 37 ℃에서 1 시간 동안 효소 Dpn1을 처리하였고, DNA 외의 PCR 산물을 제거하기 위해 PCR purification을 진행하였다. 이렇게 생성된 pET28b-EGFP와 RBD insert를 ligation하기 위해 T4 DNA polymerase를 첨가하고 상온에서 2 분 30 초 동안 반응시킨 후 얼음에서 10 분 동안 방치하여 벡터와 insert 사이의 안정적인 수소결합을 유도하였다. 그리고 클로닝 과정을 통해 얻어진 DNA 용액 4 μl를 이용하여 형질전환시켰다. 형질전환 과정은 다음과 같다. competent E.coli TOP10 용액 100 μl에 DNA 용액 4 μl를 첨가하고 얼음에서 30 분 동안 반응시킨 후에 42 ℃에서 45 초간 열처리하였다. 그리고 상기 반응액에 LB(Luria-Bertani) 액체배지 900 μl를 첨가하고 37 ℃에서 1 시간 동안 배양 후 13,000 rpm에서 10 분 동안 원심분리하여 세포를 수집하였다. 수집된 세포 용액(0.1 ml)은 카나마이신이 첨가된 LB 고체배지에 도말하고 37 ℃에서 배양한 후 형성된 콜로니 중 하나를 0.1 % 카나마이신이 첨가된 10 ml의 LB 액체배지에 접종하고 37 ℃에서 18 시간 배양하였다. 그리고 플라스미드 prep을 통해 플라스미드를 얻었다. 획득된 플라스미드의 시퀸스는 업체(바이오닉스, 한국)를 통해 확인하였다. 상기 과정에 사용된 유전자와 primer의 서열, 자세한 조성과 반응 조건은 다음과 같다.More specifically, the initial PCR was performed using primers designed complementary to both ends of the pET28b-EGFP and RBD insert 15bp. And after PCR was completed, the enzyme Dpn1 was treated at 37 °C for 1 hour to prevent self-ligation, and PCR purification was performed to remove PCR products other than DNA. To ligate the thus-produced pET28b-EGFP and RBD insert, T4 DNA polymerase was added, reacted at room temperature for 2 minutes and 30 seconds, and then left on ice for 10 minutes to induce stable hydrogen bonding between the vector and the insert. Then, it was transformed using 4 μl of the DNA solution obtained through the cloning process. The transformation process is as follows. 4 μl of DNA solution was added to 100 μl of competent E. coli TOP10 solution, and after reaction on ice for 30 minutes, heat treatment was performed at 42°C for 45 seconds. Then, 900 μl of LB (Luria-Bertani) medium was added to the reaction solution, incubated at 37° C. for 1 hour, and then centrifuged at 13,000 rpm for 10 minutes to collect cells. The collected cell solution (0.1 ml) was smeared on LB solid medium to which kanamycin was added, and one of the colonies formed after culturing at 37 ° C was inoculated into 10 ml of LB liquid medium containing 0.1% kanamycin and inoculated at 37 ° C for 18 hours. cultured. And a plasmid was obtained through plasmid prep. The sequence of the obtained plasmid was confirmed by the company (Bionics, Korea). The sequence, detailed composition and reaction conditions of the gene and primer used in the above process are as follows.
- 벡터(pET28b-EGFP, 6045bp)- vector (pET28b-EGFP, 6045bp)
① 전방 프라이머(Forward primer)① Forward primer
5’-GCTACCGCCTCCACCGTGGTG-3’ (서열번호 159)5'-GCTACCGCCTCCACCGTGGTG-3' (SEQ ID NO: 159)
② 후방 프라이머(Backward primer)② Backward primer
5’-TGAGATCCGGCTGCTAACAAAGCCCGAAA-3’ (서열번호 160)5'-TGAGATCCGGCTGCTAACAAAGCCCGAAA-3' (SEQ ID NO: 160)
- RBD 인설트- RBD Insult
① 전방 프라이머① Front primer
5’-AGCAGCCGGATCTCACAGTGGGCGTTCGCCCCATCCAT-3’ (서열번호 161)5'-AGCAGCCGGATCTCACAGTGGGCGTTCGCCCCATCCAT-3' (SEQ ID NO: 161)
② 후방 프라이머② Rear primer
5’-GGTGGAGGCGGTAGCAAAAACATTATTAATACTTCCATACTTAACCTGCGCTACGAGTCTAA-3’ (서열번호 162)5'-GGTGGAGGCGGTAGCAAAAACATTATTAATACTTCCATACTTAACCTGCGCTACGAGTCTAA-3' (SEQ ID NO: 162)
- PCR 조성, 벡터의 PCR 조건, 인설트의 PCR 조건, Dpn1 처리조건, 및 T4 DNA 폴리머레이즈 처리는 실시예 1과 상동.- PCR composition, vector PCR conditions, insult PCR conditions, Dpn1 treatment conditions, and T4 DNA polymerase treatment are the same as in Example 1.
RBD'는 site-directed mutagenesis를 이용한 방법으로 클로닝하였다. 기존 서열을 희망서열로 변경 후 서열 양 옆으로 15bp씩을 포함하여 primer를 디자인하고 backward primer는 forward primer를 상보적으로 reverse시킨다. RBD와 RBD'는 3 개의 아미노산 서열이 차이가 나기 때문에 상기 방법을 3 번 반복하여 RBD'을 제작하였다. PCR 완료 후에는 37 ℃에서 1 시간 동안 효소 Dpn1을 처리하였고 이후 형질전환하고 시퀀싱하여 염기서열을 확인하였다. 상기 과정에 사용된 유전자와 primer의 서열, 자세한 조성과 반응 조건은 다음과 같다.RBD' was cloned using site-directed mutagenesis. After changing the existing sequence to the desired sequence, design a primer including 15 bp each on each side of the sequence, and the backward primer reverses the forward primer complementaryly. Since RBD and RBD' differ in three amino acid sequences, the above method was repeated 3 times to prepare RBD'. After PCR was completed, the enzyme Dpn1 was treated at 37° C. for 1 hour, and then the nucleotide sequence was confirmed by transformation and sequencing. The sequence, detailed composition and reaction conditions of the gene and primer used in the above process are as follows.
- EGFP-RBD’ SDM1- EGFP-RBD’ SDM1
① 전방 프라이머① Front primer
5’-TATGAATTTCGTGCCACGGTAAAGACTGCTGTT -3’ (서열번호 163)5'-TATGAATTTCGTGCCACGGTAAAGACTGCTGTT -3' (SEQ ID NO: 163)
② 후방 프라이머② Rear primer
5’-AACAGCAGTCTTTACCGTGGCACGAAATTCATA -3’ (서열번호 164)5'-AACAGCAGTCTTTTACCGTGGCACGAAATTCATA -3' (SEQ ID NO: 164)
- EGFP-RBD’ SDM2- EGFP-RBD’ SDM2
① 전방 프라이머① Front primer
5’-TTCGTCGTCATTACAGACCCTCTTGGGCCCTT -3’ (서열번호 165)5'-TTCGTCGTCATTACAGACCCTCTTGGGCCCTT -3' (SEQ ID NO: 165)
② 후방 프라이머② Rear primer
5’-AAGGGCCCAAGAGGGTCTGTAATGACGACGAA -3’ (서열번호 166)5'-AAGGGCCCAAGAGGGTCTGTAATGACGACGAA -3' (SEQ ID NO: 166)
- EGFP-RBD’ SDM3- EGFP-RBD’ SDM3
① 전방 프라이머① Front primer
5’-CACATTGTTTACGTCCACGTATTTGTTTGGGTC -3’ (서열번호 167)5'-CACATTGTTTACGTCCACGTATTTGTTTGGGTC -3' (SEQ ID NO: 167)
② 후방 프라이머② Rear primer
5’- GACCCAAACAAA ACGTGGACGTAAACAATGTG -3’ (서열번호 168)5'-GACCCAAACAAA ACGTGGACGTAAACAATGTG -3' (SEQ ID NO: 168)
- PCR 조성 : 전방 프라이머 1 μL, 후방 프라이머 1 μL, 벡터 1 μL, dNTP 4 μL, 10X reaction buffer 5 μL, Pfu DNA polymerase 0.5 μL, 증류수 37.5 μL- PCR composition:
- PCR 조건- PCR conditions
① 최초 변성(95℃, 3분)① Initial denaturation (95℃, 3 minutes)
② 30회 반복: 변성(95℃, 3초) → 결합(58℃, 3초) → 연장(72℃, 6분)②
③ 마지막 연장(72℃, 3분)③ Last extension (72℃, 3 minutes)
- Dpn1 처리조건(37℃, 1시간): 5 μL(10X reaction buffer4) + 44 μL(벡터 또는 insert의 PCR 산물) + 1 μL(Dpn1용액)- Dpn1 treatment conditions (37℃, 1 hour): 5 μL (10X reaction buffer4) + 44 μL (PCR product of vector or insert) + 1 μL (Dpn1 solution)
RBD 외의 보톡스 미니어처 생산에서 Aurein1.2와 A2, A2'에 해당하는 각각의 서열은 비교적 짧기 때문에 T4 DNA polymerase를 이용한 방법이 아닌 site-directed mutagenesis 방법으로 forward primer의 앞 말단에 희망서열이 삽입되도록 디자인하여 클로닝하였다. 상기 site-directed mutagenesis 방법으로 EGFP의 C말단에 A2가 삽입되도록 하였고, EGFP와 A2사이에 Aurein1.2가 삽입되도록 플라스미드를 디자인하였다. T4 DNA polymerase를 이용해 클로닝한 EGFP-RBD도 EGFP와 RBD 사이에 Aurein1.2를 삽입하였다. PCR 완료 후 37 ℃에서 1 시간 동안 효소 Dpn1을 처리하였고 DNA 외의 PCR 산물을 제거하기 위해 PCR purification을 진행하였다. 이후 DNA들의 ligation을 위해 ligase와 kinase를 넣고 25 ℃에서 2 시간 동안 반응시킨 후 형질전환 하고 시퀀싱하여 유전자 서열을 확인하였다. 상기 과정에서 사용된 각각의 primer 염기서열과 반응조건은 다음과 같다.In Botox miniature production other than RBD, since each sequence corresponding to Aurein 1.2, A2, and A2' is relatively short, it is designed so that the desired sequence is inserted into the front end of the forward primer by site-directed mutagenesis rather than by T4 DNA polymerase. and cloned. A2 was inserted into the C-terminus of EGFP by the site-directed mutagenesis method, and a plasmid was designed so that Aurein1.2 was inserted between EGFP and A2. EGFP-RBD cloned using T4 DNA polymerase also inserted Aurein1.2 between EGFP and RBD. After PCR was completed, the enzyme Dpn1 was treated at 37 °C for 1 hour, and PCR purification was performed to remove PCR products other than DNA. Then, for ligation of DNA, ligase and kinase were added and reacted at 25 °C for 2 hours, then transformed and sequenced to confirm the gene sequence. Each primer base sequence and reaction conditions used in the above process are as follows.
- EGFP-A2- EGFP-A2
① 전방 프라이머① Front primer
5’-GGTGGAGGCGGTAGCAGAGGGAATGTAATGACGACGAACATATACCTTAACAGCAGT5'-GGTGGAGGCGGTAGCAGAGGGAATGTAATGACGACGAACATATAACCTTAACAGCAGT
TGAGATCCGGCTGCTAACAAAGCCCGAAA-3’ (서열번호 169)TGAGATCCGGCTGCTAACAAAGCCCGAAA-3' (SEQ ID NO: 169)
② 후방 프라이머② Rear primer
5’-GTGGTGGTGGTGGTGGTGCTCG-3’ (서열번호 170)5'-GTGGTGGTGGTGGTGGTGCTCG-3' (SEQ ID NO: 170)
- EGFP-Aurein1.2-A2-EGFP-Aurein1.2-A2
① 전방 프라이머① Front primer
5’-GCTACCGCCTCCACCGGCCTGTTTGACATCATCAAAAAAATCGCCGAAAGCTTTGCTAC5’-GCTACCGCCTCCACCGGCCTGTTTGACATCATCAAAAAAATCGCCGAAAGCTTTGCTAC
CGCCTCCACCGTGGTG -3’ (서열번호 171)CGCCTCCACCGTGGTG -3' (SEQ ID NO: 171)
② 후방 프라이머② Rear primer
5’- AGAGGGAATGTAATGACGACGAACATATACCTTAACAG -3’ (서열번호 172)5'-AGAGGGAATGTAATGACGACGAACATATAACCTTAACAG -3' (SEQ ID NO: 172)
- EGFP-Aurein1.2-RBD-EGFP-Aurein1.2-RBD
① 전방 프라이머① Front primer
5’-GCTACCGCCTCCACCGGCCTGTTTGACATCATCAAAAAAATCGCCGAAAGCTTTGCTAC5’-GCTACCGCCTCCACCGGCCTGTTTGACATCATCAAAAAAATCGCCGAAAGCTTTGCTAC
CGCCTCCACCGTGGTG -3’ (서열번호 173)CGCCTCCACCGTGGTG -3' (SEQ ID NO: 173)
② 후방 프라이머② Rear primer
5’- GACTCGTAGCGCAGGTTAAGTATGGAAGTATTAATAATGTTTTT -3’ (서열번호 174)5'-GACTCGTAGCGCAGGTTAAGTATGGAAGTATTAATAATGTTTTT -3' (SEQ ID NO: 174)
- PCR 조성 : 전방 프라이머 1 μL, 후방 프라이머 1 μL, 벡터 1 μL, dNTP 4 μL, 10X reaction buffer 5 μL, Pfu DNA polymerase 0.5 μL, 증류수 37.5 μL- PCR composition:
- PCR 조건- PCR conditions
① 최초 변성(95℃, 3분)① Initial denaturation (95℃, 3 minutes)
② 30회 반복: 변성(95℃, 3초) → 결합(61℃, 3초) → 연장(72℃, 6분)②
③ 마지막 연장(72℃, 3분)③ Last extension (72℃, 3 minutes)
- Dpn1 처리조건(37℃, 1시간): 5 μL(10X reaction buffer4) + 44 μL(벡터 또는 insert의 PCR 산물) + 1 μL(Dpn1용액)- Dpn1 treatment conditions (37℃, 1 hour): 5 μL (10X reaction buffer4) + 44 μL (PCR product of vector or insert) + 1 μL (Dpn1 solution)
- Ligase와 kinase 처리조건(25℃, 2시간): 5 μL(10X ligase reaction buffer + ATP) + 5 μL(10X kinase reaction buffer) + 10 μL(벡터 또는 insert의 PCR 산물) + 0.5 μL(ligase용액) + 1 μL(kinase용액) + 28.5 μL(증류수)- Ligase and kinase treatment conditions (25℃, 2 hours): 5 μL (10X ligase reaction buffer + ATP) + 5 μL (10X kinase reaction buffer) + 10 μL (PCR product of vector or insert) + 0.5 μL (ligase solution) ) + 1 μL (kinase solution) + 28.5 μL (distilled water)
※ PCR 조성 및 Dpn1 처리조건은 상기 플라스미드(1) 제작방법과 상동※ PCR composition and Dpn1 treatment conditions are the same as the above plasmid (1) production method
※ Dpn1, 10X reaction buffer, 10X reaction buffer4, 10X reaction buffer2.1, Pfu DNA polymerase T4 DNA polymerase, dNTP(2.5mM)의 구입처: ELPIS BIOTECH, 한국※ Where to purchase Dpn1, 10X reaction buffer, 10X reaction buffer4, 10X reaction buffer2.1, Pfu DNA polymerase T4 DNA polymerase, dNTP (2.5mM): ELPIS BIOTECH, Korea
※ PCR 산물의 정제법 및 플라스미드의 제조법: Dokdo-prep Gel extraction kit spintype 200 (ELPIS)에 내장된 프로토콜에 따름※ PCR product purification and plasmid preparation: Follow the protocol built in Dokdo-prep Gel extraction kit spintype 200 (ELPIS)
실시예Example 6. E. 6. E. coli를coli 이용한 뉴런 표시용 재조합 Recombination for neuron display using 녹색형광단백질green fluorescent protein 생산 및 정제 production and purification
실시예 5에서 제조된 플라스미드를 E. coli에 형질 도입하여 재조합 단백질을 생산하기 위해 발현 정도를 확인하였다. 플라스미드를 E. coli BL21(DE3)에 형질전환시키고 콜로니 하나를 선택하여 0.1 % 카나마이신이 첨가된 10 mL LB 액체배지에 넣어 37℃에서 18시간 배양하였다. 그리고 배양액 6 mL를 1 % 카나마이신이 첨가된 600 mL의 LB 액체배지에 재접종하고 200 rpm, 37 ℃에서 배양하고, 600 nm에서 광학밀도 값이 0.5에서 1 사이가 되었을 때, 0.1 mM의 IPTG를 첨가하고 150 rpm, 16 ℃에서 20 시간 동안 재조합 단백질의 발현을 유도하였다. 발현이 끝나면 8,000 rpm에서 5 분 동안 원심분리하여 세포를 수집하고, 수집된 세포는 10 mL의 PBS로 재현탁시키고 초음파 처리를 통해 세포를 파쇄하였다. 그리고 파쇄된 세포액을 12000 rpm에서 20 분 동안 원심분리하여 수용성 단백질과 불용성 단백질로 분리하고, 분리된 수용성 단백질 중 원하는 단백질만 선별하기 위해 니켈 수지를 이용하여 정제하였다. 보다 자세하게는, 1 mL의 니켈수지를 25 mL 컬럼에 넣고 125 mL의 PBS로 세척한 후 분리된 수용성 단백질 용액을 넣고 4 ℃에서 1 시간 동안 회전시키면서 부착되게 처리해준다. 부착이 끝나면 PBS와 워싱버퍼를 각각 125 mL씩 컬럼에 흘려주고 일루션 버퍼를 1 mL씩 흘려 용출된 단백질을 수집하였다. 실험 시 단계별로 샘플링하여 6X 로딩버퍼와 섞어 SDS-PAGE로 분석하였다. 상기 과정에서 사용된 버퍼 조성은 다음과 같다.By transducing the plasmid prepared in Example 5 into E. coli, the expression level was confirmed to produce a recombinant protein. The plasmid was transformed into E. coli BL21(DE3), and one colony was selected and placed in 10 mL LB broth containing 0.1% kanamycin and incubated at 37°C for 18 hours. Then, 6 mL of the culture medium was re-inoculated into 600 mL of LB broth containing 1% kanamycin, incubated at 200 rpm, 37 ° C, and when the optical density value at 600 nm was between 0.5 and 1, 0.1 mM IPTG was added. After addition, expression of the recombinant protein was induced at 150 rpm and 16° C. for 20 hours. After expression, the cells were collected by centrifugation at 8,000 rpm for 5 minutes, and the collected cells were resuspended in 10 mL of PBS and disrupted by sonication. Then, the lysed cell solution was centrifuged at 12000 rpm for 20 minutes to separate a soluble protein and an insoluble protein, and purified using a nickel resin to select only a desired protein from the separated soluble protein. In more detail, 1 mL of nickel resin is placed in a 25 mL column, washed with 125 mL of PBS, and the separated aqueous protein solution is added and treated to adhere while rotating at 4 °C for 1 hour. When the attachment was finished, 125 mL each of PBS and washing buffer was flowed into the column, and 1 mL of elution buffer was flowed to collect the eluted protein. Samples were sampled step by step during the experiment, mixed with 6X loading buffer, and analyzed by SDS-PAGE. The buffer composition used in the above process is as follows.
- PBS : NaCl 8 g/L + KCl 200 mg/L + Na2HPO4 1.44 g/L + KH2HPO4 240 mg [pH 7.4]-PBS : NaCl 8 g/L +
- 워싱 버퍼: NaCl 8 g/L + KCl 200 mg/L + Na2HPO4 1.44 g/L + KH2HPO4 240 mg + 50 mM imidazole- Washing buffer: NaCl 8 g/L +
- 일루션 버퍼: NaCl 8 g/L + KCl 200 mg/L + Na2HPO4 1.44 g/L + KH2HPO4 240 mg + 500 mM imidazole- Elution buffer: NaCl 8 g/L +
- 로딩버퍼: 0.375 M Tris [pH 6.8] + 12% SDS + 60% glycerol + 0.6 M DTT + 0.06% bromophenol blue- Loading buffer: 0.375 M Tris [pH 6.8] + 12% SDS + 60% glycerol + 0.6 M DTT + 0.06% bromophenol blue
SDS-PAGE를 통해 단백질의 발현을 분석한 결과 16 ℃, IPTG 0.1 mM의 조건에서 수용성으로 발현된 것을 확인하였다(도 9A). 발현된 재조합 녹색형광단백질과 SV2C의 결합력을 평가하기 위해 BL21(DE3) 대장균을 이용하여 수용성으로 발현된 단백질을 정제하고자 하였다. 형광단백질과 융합된 재조합 녹색형광단백질과 SV2C은 히스티딘 태그(His6-tag)를 이용해 니켈 bead로 정제하였고, 수용체는 GST(glutathione S-transferase) 태그를 이용해 glutathione bead로 정제하였으며 SDS-PAGE를 통해 분석하였다(도 9B).As a result of analyzing the expression of the protein through SDS-PAGE, it was confirmed that the protein was expressed in water at 16 °C and 0.1 mM IPTG (FIG. 9A). In order to evaluate the binding force between the expressed recombinant green fluorescent protein and SV2C, it was attempted to purify the expressed protein in water using BL21(DE3) E. coli. Recombinant green fluorescent protein and SV2C fused with fluorescent protein were purified with nickel beads using histidine tag (His6-tag), and the receptor was purified with glutathione beads using GST (glutathione S-transferase) tag. (FIG. 9B).
실시예Example 7. 7. GSTGST pull assay를 이용한 뉴런 표시용 재조합 Recombination for neuron display using pull assay 녹색형광단백질과green fluorescent protein 뉴런 수용체( neuronal receptors ( SV2CSV2C )의 결합력 확인) check the binding force of
실시예 6에서 발현/정제된 재조합 녹색형광단백질과 SV2C 간의 결합력을 확인하기 위해 GST pull down을 진행하였다. 정제된 SV2C를 glutathione bead에 결합시킨 후 재조합 녹색형광단백질과 상기 SV2C와 결합된 glutathione bead를 반응시켰다. 결합하지 못한 단백질을 제거하기 위해 bead를 세척한 후 SDS-PAGE 내에서 전기영동하여 확인하였다(도 10). 이 때 세척에 사용한 버퍼는 두 가지 pH(pH 7.4와 5.8)로 나누어 사용하였다. SDS-PAGE 결과에 따르면, pH 7.4와 pH 5.8 워싱버퍼 사용 간의 차이가 없는 것으로 보아 pH에 의존적이지 않게 결합한다는 것을 알 수 있었으며, 각각 RBD 보다는 RBD'이, A2 보다는 A2'이, A2와 A2' 보다는 RBD와 RBD'이, SV2C와의 결합력이 좋다는 것을 확인할 수 있었다. 따라서 본 실시예를 통해 실시예 6에서 제조된 뉴런 표시용 재조합 녹색형광단백질과 SV2C 간의 결합력이 존재하는 것을 확인할 수 있었다.In order to confirm the binding force between the recombinant green fluorescent protein expressed/purified in Example 6 and SV2C, GST pull down was performed. After binding the purified SV2C to the glutathione bead, the recombinant green fluorescent protein was reacted with the glutathione bead bound to the SV2C. After washing the beads to remove the unbound protein, it was confirmed by electrophoresis in SDS-PAGE (FIG. 10). At this time, the buffer used for washing was divided into two pHs (pH 7.4 and 5.8). According to the SDS-PAGE results, there was no difference between the use of pH 7.4 and pH 5.8 washing buffers, indicating that the binding was not dependent on pH, and RBD' rather than RBD, A2' rather than A2, A2 and A2', respectively. Rather, it was confirmed that RBD and RBD' had good binding ability with SV2C. Therefore, it was confirmed that the binding force between the recombinant green fluorescent protein for neuron display and SV2C prepared in Example 6 was present through this Example.
실시예Example 8. 8. 공초점confocal 레이저 형광 현미경 이미지 획득 Laser Fluorescence Microscopy Image Acquisition
콜라겐 용액과 poly-D-lysine 용액에 각각 12 시간씩 코팅된 커버글라스를 세포 배양용 12 well plate에 넣고 PC12 cell(1.0 X 105 cell/well)을 분주하여 37 ℃ 5 % CO2 배양기에서 하루 동안 세포를 배양하였다. 그 후 100 ng/mL nerve growth factor(NGF)를 처리하여 5일 동안 분화를 유도하였다. 분화가 확인되면 실시예 6에서 발현/정제된 재조합 녹색형광단백질을 첨가하고 15 분 동안 37 ℃ 5 % CO2 배양기에서 배양해준 후, 엔도솜과 핵(Hoechst 33258), 그리고 세포막(WGA-AF647)을 차례로 염색하였다. 염색이 끝나면 커버글라스만 핀셋으로 꺼내 슬라이드글라스 위에 덮고 움직이지 않게 고정시키고, 상기 방법으로 만들어진 샘플을 공초점 레이저 형광 현미경(Leica TCS SP8 HyVolution confocal microscope)으로 확인하였다. 그 결과는 도 11에 나타내었다.A cover glass coated with collagen solution and poly-D-lysine solution for 12 hours each was placed in a 12 well plate for cell culture, and PC12 cells (1.0 X 10 5 cell/well) were dispensed and incubated at 37 ° C. 5% CO 2 for one day. cells were cultured during Thereafter, differentiation was induced for 5 days by treatment with 100 ng/mL nerve growth factor (NGF). When differentiation is confirmed, the recombinant green fluorescent protein expressed/purified in Example 6 is added, and incubated in an incubator at 37 ° C. 5% CO 2 for 15 minutes, endosomes, nuclei (Hoechst 33258), and cell membrane (WGA-AF647) were sequentially stained. After staining, only the cover glass was taken out with tweezers, covered with tweezers, and fixed immovably, and the sample prepared by the above method was checked with a Leica TCS SP8 HyVolution confocal microscope. The results are shown in FIG. 11 .
도 11에서 확인할 수 있듯이 EGFP-A2, EGFP-A2', EGFP-RBD, EGFP-RBD' 모두 세포 내에서도 EGFP에 해당하는 녹색형광 신호가 관찰되었기 때문에, 실시예 6에서 제조된 뉴런 표시용 재조합 녹색형광단백질은 신경세포를 표시하는 역할을 잘 수행할 수 있다는 것을 확인하였다. As can be seen in FIG. 11 , since green fluorescence signals corresponding to EGFP were observed in all cells of EGFP-A2, EGFP-A2', EGFP-RBD, and EGFP-RBD', the recombinant green fluorescence for displaying neurons prepared in Example 6 It was confirmed that the protein can perform the role of marking nerve cells well.
- 배양배지: RPMI-1640 + 5% fetal bovine serum + 10% horse serum + 1% antibiotics- Culture medium: RPMI-1640 + 5% fetal bovine serum + 10% horse serum + 1% antibiotics
- 콜라겐 용액 : 50 μg/mL collagen in 20 mM acetic acid- Collagen solution: 50 μg/mL collagen in 20 mM acetic acid
- Poly-D-lysine 용액 : 0.1 mg/mL Poly-D-Lysine- Poly-D-lysine solution: 0.1 mg/mL Poly-D-Lysine
상기 결과들을 통하여, 본 발명의 EEP와 BoNT/A-A2 또는 RBD를 신경-활성 물질에 다양한 형태로 결합시킴으로써, 신경-활성 물질 재조합 단백질이 뉴런을 특이적으로 인식하고 뉴런의 세포질로 탈출하여 활성을 나타내는 것을 확인할 수 있었으며, 이를 통하여, 다양한 신경-활성 물질에 EEP와 A2 또는 RBD를 결합시킴으로써 뉴론 특이적으로 다양한 약물 전달, 형광 신호를 통한 뉴런 관찰 등 다양한 분야에 적용할 수 있다는 것을 확인할 수 있었다. Through the above results, by binding the EEP of the present invention and BoNT/A-A2 or RBD to neuro-active substances in various forms, the neuro-active substance recombinant protein specifically recognizes neurons and escapes into the cytoplasm of neurons and is active was confirmed, and through this, it was confirmed that by binding EEP and A2 or RBD to various neuro-active substances, it could be applied to various fields such as neuron-specific drug delivery and neuron observation through fluorescence signals. .
<110> Research and Business Foundation SungKyunKwan University <120> A composition comprising neuro-targeted proteins <130> MP19-178 <150> KR 10-2018-0101615 <151> 2018-08-28 <160> 174 <170> KoPatentIn 3.0 <210> 1 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 <400> 1 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Lys Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Lys Gly Tyr Asn Lys 435 440 445 <210> 2 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype <400> 2 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Val Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Lys Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Lys Gly Tyr Asn Lys 435 440 445 <210> 3 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 3 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 4 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype <400> 4 Met Gln Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Val Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Lys Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Lys Gly Tyr Asn Lys 435 440 445 <210> 5 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A5 subtype <400> 5 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Glu Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Glu His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Glu Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 6 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 6 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Ile Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 7 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A5 subtype <400> 7 Met Leu Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Glu Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Glu His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Glu Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 8 <211> 444 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A3 subtype <400> 8 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Arg Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Glu Gly Val Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Ile Lys Leu Phe Asp 85 90 95 Arg Ile Tyr Ser Thr Gly Leu Gly Arg Met Leu Leu Ser Phe Ile Val 100 105 110 Lys Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Glu Pro Gly Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Thr Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Phe Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Thr Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Ala His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Leu Lys Val Lys Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly Asn Asp Thr Asn 260 265 270 Phe Ile Asp Ser Leu Trp Gln Lys Lys Phe Ser Arg Asp Ala Tyr Asp 275 280 285 Asn Leu Gln Asn Ile Ala Arg Ile Leu Asn Glu Ala Lys Thr Ile Val 290 295 300 Gly Thr Thr Thr Pro Leu Gln Tyr Met Lys Asn Ile Phe Ile Arg Lys 305 310 315 320 Tyr Phe Leu Ser Glu Asp Ala Ser Gly Lys Ile Ser Val Asn Lys Pro 325 330 335 Ala Phe Lys Glu Phe Tyr Arg Val Leu Thr Arg Gly Phe Thr Glu Leu 340 345 350 Glu Phe Val Asn Pro Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Asn Glu Gly Phe Asn Leu Glu Gly Ala Asn Ser Asn Gly Gln 385 390 395 400 Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu Lys Asn Phe Thr 405 410 415 Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg Gly Ile Ile Pro 420 425 430 Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 <210> 9 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 9 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Leu Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 10 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 10 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Ile Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 11 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A <400> 11 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Ile Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Asn Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Ile Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Glu Val Ala Ser Ile Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Arg Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Met Asn Ile Val Pro Glu Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 12 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 12 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Thr Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Lys Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asn Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 13 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A4 subtype <400> 13 Met Pro Leu Val Asn Gln Gln Ile Asn Tyr Tyr Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Lys Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Val Trp Val Ile Pro Glu Arg 35 40 45 Asp Ile Phe Thr Asn Pro Glu Glu Val Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Ile Ser Tyr Tyr Asp Ser Ala Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Ile Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Ile Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Gly Lys Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Ile Ile Gln Leu Asp Asp Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Ala Ile Ile Gly Pro Ser Ala Asn Ile 145 150 155 160 Ile Glu Ser Gln Cys Ser Ser Phe Arg Asp Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Val Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Gln Asp Pro Ala Val Ala Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Thr Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ala Gly Leu 245 250 255 Glu Val Ser Leu Glu Glu Leu Ile Thr Phe Gly Gly Asn Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Lys Lys Glu Phe Ser Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Ala Thr Gly Lys Phe Leu Val Asp Arg Leu 325 330 335 Lys Phe Asp Glu Leu Tyr Lys Leu Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Asp Val Asn Tyr 370 375 380 Thr Ile His Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Ile Glu Ile Asn Asn Lys Asn Phe Asp Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 14 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 14 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Glu Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 15 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype <400> 15 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Val 20 25 30 Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg Asp 35 40 45 Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu Ala 50 55 60 Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr Asp 65 70 75 80 Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu Arg 85 90 95 Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val Arg 100 105 110 Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys Val 115 120 125 Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr Arg 130 135 140 Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile Ile 145 150 155 160 Gln Phe Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn Leu Thr Arg 165 170 175 Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe Thr 180 185 190 Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu Gly 195 200 205 Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu Leu 210 215 220 Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn Arg 225 230 235 240 Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu Glu 245 250 255 Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys Phe 260 265 270 Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn Lys 275 280 285 Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val Gly 290 295 300 Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys Tyr 305 310 315 320 Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu Lys 325 330 335 Phe Asp Lys Leu Tyr Lys Met Lys Thr Glu Ile Tyr Thr Glu Asp Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Lys Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 16 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 16 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asp Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Phe Gln Thr Leu Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Thr Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asn Lys Leu Tyr Lys Ser Leu 340 345 350 Met Leu Gly Phe Thr Glu Ile Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Gly Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 17 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B-B1 subtype <400> 17 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 18 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 18 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asp Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Gln Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Thr Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asn Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Ile Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Gly Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 19 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 19 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Met Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asn Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 20 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 20 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asp Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Gln Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Thr Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asn Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Gly Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 21 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 21 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Arg 435 440 <210> 22 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 22 Met Ser Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asp Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asn Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 23 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 23 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asp Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 24 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B-B8 subtype <400> 24 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Gly Glu Glu Arg Lys Glu Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asp Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Gly Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Arg 435 440 <210> 25 <211> 439 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B-B1 subtype <400> 25 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Ile Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Met Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Leu Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Trp Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Ala Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Ile Val Asn Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Phe Val Glu Asp Ser Glu Gly Lys Tyr 325 330 335 Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu Met Phe 340 345 350 Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys Thr Arg 355 360 365 Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys Asn Leu 370 375 380 Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile Ser Asp 385 390 395 400 Lys Asn Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile Asn Lys 405 410 415 Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Ile Tyr Lys Ile 420 425 430 Gln Met Cys Lys Ser Val Lys 435 <210> 26 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B-B8 subtype <400> 26 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Gly Glu Glu Arg Lys Glu Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Gly Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asp Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Gly Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Arg 435 440 <210> 27 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 27 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Trp Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Ala Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Ile Val Asn Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 28 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 28 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Ser Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 29 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/C1 <400> 29 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Thr Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asn Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Val 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Asp Lys Asp Pro Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ser Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Thr Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Thr Val Asn Arg Asn Lys Phe Val Glu Leu Tyr Asn 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 30 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/C1 <400> 30 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Ile Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asp Ser Asn Pro Asn Leu Lys Lys Pro Pro Arg Val 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Glu Lys Asp Thr Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ala Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Arg Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Glu 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Ala Val Asp Arg Asn Lys Phe Ala Glu Leu Tyr Lys 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 31 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/C1 <400> 31 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Ile Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asp Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Val 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Glu Lys Asp Thr Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ala Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asn 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Thr Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Arg Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Lys Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Gly Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Ala Val Asp Arg Asn Lys Phe Ala Glu Leu Tyr Lys 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 32 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/C1 <400> 32 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Ile Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asp Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Xaa 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Glu Lys Asp Thr Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ala Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asn 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Arg Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Lys Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Ala Val Asp Arg Asn Lys Phe Ala Glu Leu Tyr Lys 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Ala Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 33 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/CD <400> 33 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Ile Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asp Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Val 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Glu Lys Asp Thr Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ala Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asn 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Arg Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Ala Val Asp Arg Asn Lys Phe Ala Glu Leu Tyr Lys 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 34 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/CD <400> 34 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Ile Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asp Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Val 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Glu Lys Asp Thr Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ala Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asn 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Arg Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Lys Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Ala Val Asp Arg Asn Lys Phe Ala Glu Leu Tyr Lys 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 35 <211> 442 <212> PRT <213> Artificial Sequence <220> <223> BoNT/D <400> 35 Met Thr Trp Pro Val Lys Asp Phe Asn Tyr Ser Asp Pro Val Asn Asp 1 5 10 15 Asn Asp Ile Leu Tyr Leu Arg Ile Pro Gln Asn Lys Leu Ile Thr Thr 20 25 30 Pro Val Lys Ala Phe Met Ile Thr Gln Asn Ile Trp Val Ile Pro Glu 35 40 45 Arg Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro 50 55 60 Thr Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp 65 70 75 80 Glu Gln Lys Asp Thr Phe Leu Lys Gly Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Glu Arg Asp Ile Gly Lys Lys Leu Ile Asn Tyr Leu Val Val 100 105 110 Gly Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp 115 120 125 Phe Thr Arg His Thr Thr Asn Ile Ala Val Glu Lys Phe Glu Asn Gly 130 135 140 Ser Trp Lys Val Thr Asn Ile Ile Thr Pro Ser Val Leu Ile Phe Gly 145 150 155 160 Pro Leu Pro Asn Ile Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly 165 170 175 Gln Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu 180 185 190 Lys Val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr Ser Asn 195 200 205 Gln Ser Ser Ala Val Leu Gly Lys Ser Ile Phe Cys Met Asp Pro Val 210 215 220 Ile Ala Leu Met His Glu Leu Thr His Ser Leu His Gln Leu Tyr Gly 225 230 235 240 Ile Asn Ile Pro Ser Asp Lys Arg Ile Arg Pro Gln Val Ser Glu Gly 245 250 255 Phe Phe Ser Gln Asp Gly Pro Asn Val Gln Phe Glu Glu Leu Tyr Thr 260 265 270 Phe Gly Gly Leu Asp Val Glu Ile Ile Pro Gln Ile Glu Arg Ser Gln 275 280 285 Leu Arg Glu Lys Ala Leu Gly His Tyr Lys Asp Ile Ala Lys Arg Leu 290 295 300 Asn Asn Ile Asn Lys Thr Ile Pro Ser Ser Trp Ile Ser Asn Ile Asp 305 310 315 320 Lys Tyr Lys Lys Ile Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn 325 330 335 Thr Gly Asn Phe Val Val Asn Ile Asp Lys Phe Asn Ser Leu Tyr Ser 340 345 350 Asp Leu Thr Asn Val Met Ser Glu Val Val Tyr Ser Ser Gln Tyr Asn 355 360 365 Val Lys Asn Arg Thr His Tyr Phe Ser Arg His Tyr Leu Pro Val Phe 370 375 380 Ala Asn Ile Leu Asp Asp Asn Ile Tyr Thr Ile Arg Asp Gly Phe Asn 385 390 395 400 Leu Thr Asn Lys Gly Phe Asn Ile Glu Asn Ser Gly Gln Asn Ile Glu 405 410 415 Arg Asn Pro Ala Leu Gln Lys Leu Ser Ser Glu Ser Val Val Asp Leu 420 425 430 Phe Thr Lys Val Cys Leu Arg Leu Thr Lys 435 440 <210> 36 <211> 442 <212> PRT <213> Artificial Sequence <220> <223> BoNT/D <400> 36 Met Thr Trp Pro Val Lys Asp Phe Asn Tyr Ser Asp Pro Val Asn Asp 1 5 10 15 Asn Asp Ile Leu Tyr Leu Arg Ile Pro Gln Asn Lys Leu Ile Thr Thr 20 25 30 Pro Val Lys Ala Phe Met Ile Thr Gln Asn Ile Trp Val Ile Pro Glu 35 40 45 Arg Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro 50 55 60 Thr Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp 65 70 75 80 Glu Gln Lys Asp Thr Phe Leu Lys Gly Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Glu Arg Asp Ile Gly Lys Lys Leu Ile Asn Tyr Leu Val Val 100 105 110 Gly Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp 115 120 125 Phe Thr Arg His Thr Thr Asn Ile Ala Val Glu Lys Phe Glu Asn Gly 130 135 140 Ser Trp Lys Val Thr Asn Ile Ile Thr Pro Ser Val Leu Ile Phe Gly 145 150 155 160 Pro Leu Pro Asn Ile Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly 165 170 175 Gln Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu 180 185 190 Lys Val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr Ser Asn 195 200 205 Gln Ser Ser Ala Val Leu Gly Lys Ser Ile Phe Cys Met Asp Pro Val 210 215 220 Ile Ala Leu Met His Glu Leu Thr His Ser Leu His Gln Leu Tyr Gly 225 230 235 240 Ile Asn Ile Pro Ser Asp Lys Arg Ile Arg Pro Gln Val Ser Glu Gly 245 250 255 Phe Phe Ser Gln Asp Gly Pro Asn Val Gln Phe Glu Glu Leu Tyr Thr 260 265 270 Phe Gly Gly Ser Asp Val Glu Ile Ile Pro Gln Ile Glu Arg Ser Gln 275 280 285 Leu Arg Glu Lys Ala Leu Asp His Tyr Lys Asp Ile Ala Lys Arg Leu 290 295 300 Asn Asn Ile Asn Lys Thr Ile Pro Ser Ser Trp Ser Ser Asn Ile Asp 305 310 315 320 Lys Tyr Lys Lys Ile Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn 325 330 335 Thr Gly Asn Phe Val Val Ser Ile Asp Lys Phe Asn Arg Leu Tyr Ser 340 345 350 Asp Leu Thr Asn Val Met Ser Glu Val Val Tyr Ser Ser Gln Tyr Asn 355 360 365 Val Lys Asn Arg Thr His Tyr Phe Ser Lys Arg Tyr Leu Pro Val Phe 370 375 380 Ala Asn Ile Leu Asp Asp Asn Ile Tyr Thr Ile Ile Asp Gly Phe Asn 385 390 395 400 Leu Thr Thr Lys Gly Phe Asn Ile Glu Asn Ser Gly Gln Asn Ile Glu 405 410 415 Arg Asn Pro Ala Leu Gln Lys Leu Ser Ser Glu Ser Val Val Asp Leu 420 425 430 Phe Thr Lys Val Cys Leu Arg Leu Thr Lys 435 440 <210> 37 <211> 442 <212> PRT <213> Artificial Sequence <220> <223> BoNT/D <400> 37 Met Thr Trp Pro Val Lys Asp Phe Asn Tyr Ser Asp Pro Val Asn Asp 1 5 10 15 Asn Asp Ile Leu Tyr Leu Arg Ile Pro Gln Asn Lys Leu Ile Thr Thr 20 25 30 Pro Val Lys Ala Phe Met Ile Thr Gln Asn Ile Trp Val Ile Pro Glu 35 40 45 Arg Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro 50 55 60 Thr Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp 65 70 75 80 Glu Gln Lys Asp Thr Phe Leu Lys Gly Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Glu Arg Asp Ile Gly Lys Lys Leu Ile Asn Tyr Leu Val Val 100 105 110 Gly Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp 115 120 125 Phe Thr Arg His Thr Thr Asn Ile Ala Val Glu Lys Phe Glu Asn Gly 130 135 140 Ser Trp Lys Val Thr Asn Ile Ile Thr Pro Ser Val Leu Ile Phe Gly 145 150 155 160 Pro Leu Pro Asn Ile Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly 165 170 175 Gln Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu 180 185 190 Lys Val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr Ser Asn 195 200 205 Gln Ser Ser Ala Val Leu Gly Lys Ser Ile Phe Cys Met Asp Pro Val 210 215 220 Ile Ala Leu Met His Glu Leu Thr His Ser Leu His Gln Leu Tyr Gly 225 230 235 240 Ile Asn Ile Pro Ser Asp Lys Arg Ile Arg Pro Gln Val Ser Glu Gly 245 250 255 Phe Phe Ser Gln Asp Gly Pro Asn Val Gln Phe Glu Glu Leu Tyr Thr 260 265 270 Phe Gly Gly Ser Asp Val Glu Ile Ile Pro Gln Ile Glu Arg Leu Gln 275 280 285 Leu Arg Glu Lys Ala Leu Gly His Tyr Lys Asp Ile Ala Lys Arg Leu 290 295 300 Asn Asn Ile Asn Lys Thr Ile Pro Ser Ser Trp Ser Ser Asn Ile Asp 305 310 315 320 Lys Tyr Lys Lys Ile Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn 325 330 335 Thr Gly Asn Phe Val Val Asn Ile Asp Lys Phe Asn Ser Leu Tyr Ser 340 345 350 Asp Leu Thr Asn Val Met Ser Glu Val Val Tyr Ser Ser Gln Tyr Asn 355 360 365 Val Lys Asn Arg Thr His Tyr Phe Ser Lys His Tyr Leu Pro Val Phe 370 375 380 Ala Asn Ile Leu Asp Asp Asn Ile Tyr Thr Ile Ile Asn Gly Phe Asn 385 390 395 400 Leu Thr Thr Lys Gly Phe Asn Ile Glu Asn Ser Gly Gln Asn Ile Glu 405 410 415 Arg Asn Pro Ala Leu Gln Lys Leu Ser Ser Glu Ser Val Val Asp Leu 420 425 430 Phe Thr Lys Val Cys Leu Arg Leu Thr Arg 435 440 <210> 38 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 38 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Asp Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Arg Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Cys Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Arg Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 39 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 39 Met Pro Thr Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asn Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Gln Glu Lys 65 70 75 80 Asp Lys Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Arg Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Gly Asp Phe Ile Ile Asn Asp 115 120 125 Ala Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Lys Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 40 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E3 subtype <400> 40 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln His Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Ile Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Thr Cys Ile Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Arg Lys Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Asn Asp Leu Asn Ile Ile Thr Val Ala Gln Tyr Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asn Asp Tyr Arg Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asp Asp Ile Leu Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Lys Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 41 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E1 subtype <400> 41 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Asp Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 42 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 42 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Lys 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Arg Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Asn Gln Ser Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Arg Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Tyr Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asn Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asn Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Thr Asn Leu Asn Pro Arg Ile Ile Thr Gln Leu Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Phe Ser Lys Gly Ile Thr 420 <210> 43 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 43 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ile Gln Asp Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 44 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 44 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln His Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Ile Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Thr Cys Ile Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Arg Lys Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Asn Asp Leu Asn Ile Ile Thr Val Ala Gln Tyr Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asn Asp Tyr Arg Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Lys Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 45 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E10 subtype <400> 45 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Lys 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asn Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Thr Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Lys Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Asn Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Gln 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asn Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asn Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Thr Asn Leu Asn Pro Arg Ile Ile Thr Gln Leu Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Phe Ser Lys Gly Ile Thr 420 <210> 46 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 46 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Asn 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asn Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Asn Asp Gln Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Val Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Arg Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Arg Asp Asp Asp Phe Ile Ile Asn Asp 115 120 125 Gly Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Thr Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Val Ser Leu Ile Asn Asn Tyr Ser Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Ile His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Asn Asn Leu Asn Ile Ile Thr Ser Ser Gln Leu Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asp Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Val Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asn Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Asn Val Asn Phe 370 375 380 Arg Gly Gln Asn Pro Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Asp Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 47 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 47 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Asp Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Gln Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 48 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 48 Met Pro Thr Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Tyr Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Asp Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Lys Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Gln Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Lys Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Tyr Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Lys Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asn Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Thr Asn Leu Asn Pro Arg Ile Ile Thr Pro Leu Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Phe Ser Lys Gly Ile Arg 420 <210> 49 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E10 subtype <400> 49 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Lys 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asn Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Thr Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Lys Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Asn Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Gln 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asn Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asn Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Thr Asn Leu Asn Pro Arg Ile Ile Lys Gln Leu Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Ser Lys Gly Ile Thr 420 <210> 50 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E11 subtype <400> 50 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Lys 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asn Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Thr Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Lys Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Asn Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asp Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Gln 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Val Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asp Ala Asn Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Lys Thr Tyr Ile 340 345 350 Gly His His Lys Tyr Phe Arg Leu Ser Asp Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Asp Gly Tyr Asn Ile Asn Thr Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Thr Asn Leu Asn Thr Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Val Val Arg Lys Ile Ile Arg Phe Cys Thr Asn Ile Phe 405 410 415 Ser Pro Lys Gly Ile Arg 420 <210> 51 <211> 425 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E1 subtype <400> 51 Met Leu Tyr Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val 1 5 10 15 Asn Asp Arg Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe 20 25 30 Tyr Lys Ser Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg 35 40 45 Asn Val Ile Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu 50 55 60 Lys Asn Gly Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp 65 70 75 80 Glu Glu Lys Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg 85 90 95 Ile Asn Asn Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys 100 105 110 Ala Asn Pro Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His 115 120 125 Ile Gly Asp Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln 130 135 140 Asp Ile Leu Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu 145 150 155 160 Phe Glu Thr Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro 165 170 175 Ser Asn His Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu 180 185 190 Tyr Ser Phe Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp 195 200 205 Pro Ala Leu Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu 210 215 220 Tyr Gly Ala Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln 225 230 235 240 Asn Pro Leu Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu 245 250 255 Thr Phe Gly Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn 260 265 270 Asp Ile Tyr Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys 275 280 285 Leu Ser Lys Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp 290 295 300 Val Phe Glu Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr 305 310 315 320 Ser Val Asn Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser 325 330 335 Phe Thr Glu Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln 340 345 350 Thr Tyr Ile Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn 355 360 365 Asp Ser Ile Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys 370 375 380 Val Asn Phe Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr 385 390 395 400 Pro Ile Thr Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys 405 410 415 Asn Ile Val Ser Val Lys Gly Ile Arg 420 425 <210> 52 <211> 425 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E10 subtype <400> 52 Met Leu Tyr Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val 1 5 10 15 Asn Asp Lys Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe 20 25 30 Tyr Lys Ser Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg 35 40 45 Asn Val Ile Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu 50 55 60 Lys Asn Gly Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn 65 70 75 80 Glu Glu Lys Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg 85 90 95 Ile Asn Asp Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys 100 105 110 Ala Asn Pro Tyr Leu Gly Asn Asp Asn Thr Pro Asn Asn Gln Phe His 115 120 125 Ile Gly Asp Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln 130 135 140 Ser Ile Leu Leu Pro Thr Val Ile Ile Met Gly Ala Glu Pro Asp Leu 145 150 155 160 Phe Glu Thr Asn Ser Ser Asn Ile Ser Leu Lys Asn Asn Tyr Met Pro 165 170 175 Ser Asn His Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu 180 185 190 Tyr Ser Phe Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp 195 200 205 Pro Ala Leu Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu 210 215 220 Tyr Gly Ala Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln 225 230 235 240 Asn Pro Leu Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu 245 250 255 Thr Phe Gly Gly Thr Asp Leu Asn Ile Ile Thr Asn Ala Gln Ser Asn 260 265 270 Asp Ile Tyr Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys 275 280 285 Leu Ser Gln Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp 290 295 300 Ile Phe Gln Glu Lys Tyr Gly Leu Asp Lys Asn Ala Ser Gly Ile Tyr 305 310 315 320 Ser Val Asn Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser 325 330 335 Phe Thr Glu Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln 340 345 350 Thr Tyr Ile Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn 355 360 365 Asn Ser Ile Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Lys 370 375 380 Val Asn Phe Arg Gly Gln Asn Thr Asn Leu Asn Pro Arg Ile Ile Thr 385 390 395 400 Gln Leu Thr Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys 405 410 415 Asn Ile Val Phe Ser Lys Gly Ile Thr 420 425 <210> 53 <211> 425 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E3 subtype <400> 53 Met Leu Tyr Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val 1 5 10 15 Asn Asp Arg Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe 20 25 30 Tyr Lys Ser Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg 35 40 45 Asn Val Ile Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu 50 55 60 Lys Asn Gly Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp 65 70 75 80 Glu Glu Lys Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg 85 90 95 Ile Asn Asn Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys 100 105 110 Ala Asn Pro Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His 115 120 125 Ile Gly Asp Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln 130 135 140 His Ile Leu Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu 145 150 155 160 Phe Glu Thr Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro 165 170 175 Ser Asn His Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu 180 185 190 Tyr Ser Phe Arg Phe Asn Asp Asn Ser Ile Asn Glu Phe Ile Gln Asp 195 200 205 Pro Ala Leu Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu 210 215 220 Tyr Gly Ala Lys Gly Ile Thr Thr Thr Cys Ile Ile Thr Gln Gln Gln 225 230 235 240 Asn Pro Leu Ile Thr Asn Arg Lys Gly Ile Asn Ile Glu Glu Phe Leu 245 250 255 Thr Phe Gly Gly Asn Asp Leu Asn Ile Ile Thr Val Ala Gln Tyr Asn 260 265 270 Asp Ile Tyr Thr Asn Leu Leu Asn Asp Tyr Arg Lys Ile Ala Ser Lys 275 280 285 Leu Ser Lys Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp 290 295 300 Ile Phe Gln Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr 305 310 315 320 Ser Val Asn Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser 325 330 335 Phe Thr Glu Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu 340 345 350 Thr Tyr Ile Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn 355 360 365 Asp Ser Ile Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys 370 375 380 Val Asn Phe Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Lys 385 390 395 400 Pro Ile Thr Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys 405 410 415 Asn Ile Val Ser Val Lys Gly Ile Arg 420 425 <210> 54 <211> 425 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 54 Met Leu Tyr Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val 1 5 10 15 Asn Asp Arg Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe 20 25 30 Tyr Lys Ser Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg 35 40 45 Asn Val Ile Gly Thr Ile Pro Gln Asp Phe Gln Pro Pro Thr Ser Leu 50 55 60 Lys Asn Gly Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn 65 70 75 80 Glu Glu Lys Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg 85 90 95 Ile Asn Asp Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys 100 105 110 Ala Asn Pro Tyr Leu Gly Asn Asp Asn Thr Pro Asp Gly Asp Phe Ile 115 120 125 Ile Asn Asp Ala Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser Gln 130 135 140 Ser Ile Leu Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu 145 150 155 160 Phe Glu Thr Asn Ser Ser Asn Ile Ser Leu Ile Asn Asn Tyr Arg Pro 165 170 175 Ser Asn His Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu 180 185 190 Tyr Ser Phe Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp 195 200 205 Pro Ala Leu Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu 210 215 220 Tyr Gly Ala Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln 225 230 235 240 Asn Ser Leu Ile Thr Asn Ile Arg Gly Ile Asn Ile Glu Glu Phe Leu 245 250 255 Thr Phe Gly Gly Asn Asp Leu Asn Ile Ile Thr Ser Ser Gln Phe Asn 260 265 270 Asp Ile Tyr Thr Asn Leu Leu Asp Asp Tyr Lys Lys Ile Ala Ser Lys 275 280 285 Leu Ser Gln Val Arg Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp 290 295 300 Val Phe Gln Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr 305 310 315 320 Ser Val Asn Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser 325 330 335 Phe Thr Glu Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu 340 345 350 Thr Tyr Ile Gly Gln Tyr Lys Tyr Phe Gln Leu Ser Asn Leu Leu Asn 355 360 365 Asp Ser Ile Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys 370 375 380 Val Asn Phe Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr 385 390 395 400 Pro Ile Thr Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys 405 410 415 Asn Ile Val Ser Val Lys Gly Ile Arg 420 425 <210> 55 <211> 421 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 55 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Pro Val Asn Asp Arg Thr 1 5 10 15 Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser Phe 20 25 30 Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile Gly 35 40 45 Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly Asp 50 55 60 Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys Asp 65 70 75 80 Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn Asn 85 90 95 Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro Tyr 100 105 110 Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp Ala 115 120 125 Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln His Ile Leu Leu 130 135 140 Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr Asn 145 150 155 160 Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His Gly 165 170 175 Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe Arg 180 185 190 Phe Asn Asp Asn Ser Ile Asn Glu Phe Ile Gln Asp Pro Ala Leu Thr 195 200 205 Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala Lys 210 215 220 Gly Ile Thr Thr Thr Cys Ile Ile Thr Gln Gln Gln Asn Pro Leu Ile 225 230 235 240 Thr Asn Arg Lys Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly Gly 245 250 255 Asn Asp Leu Asn Ile Ile Thr Val Ala Gln Tyr Asn Asp Ile Tyr Thr 260 265 270 Asn Leu Leu Asn Asp Tyr Arg Lys Ile Ala Ser Lys Leu Ser Lys Val 275 280 285 Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln Glu 290 295 300 Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn Ile 305 310 315 320 Asn Lys Phe Asp Asp Ile Leu Lys Lys Leu Tyr Ser Phe Thr Glu Phe 325 330 335 Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile Gly 340 345 350 Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile Tyr 355 360 365 Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe Arg 370 375 380 Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Lys Pro Ile Thr Gly 385 390 395 400 Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val Ser 405 410 415 Val Lys Gly Ile Arg 420 <210> 56 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 56 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln His Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Ile Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Thr Cys Ile Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Arg Lys Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Asn Asp Leu Asn Ile Ile Thr Val Ala Gln Tyr Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asn Asp Tyr Arg Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe His Asp Ile Leu Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Lys Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 57 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 57 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Gln Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Gly Asp Phe Ile Ile Asn Asp 115 120 125 Ala Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Ile Asn Asn Tyr Arg Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Ser Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Asn Asp Leu Asn Ile Ile Thr Ser Ser Gln Phe Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asp Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Gln 275 280 285 Val Arg Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Val Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Gln Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 58 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 58 Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Lys Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asp Glu Phe 115 120 125 Ser Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Leu Ser Thr Asn 130 135 140 Val Glu Ser Ser Met Leu Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Glu Ser Cys Cys Tyr Pro Val Arg Lys Leu Ile Asp Pro 165 170 175 Asp Val Val Tyr Asp Pro Ser Asn Tyr Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Glu Glu Thr Ile Glu Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ser Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Glu Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Ser Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val 420 425 430 Ile Pro Arg Lys 435 <210> 59 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 59 Met Pro Val Val Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asp Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Glu Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Glu Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Glu His Thr Pro Ile Asn Glu Phe 115 120 125 His Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Ser Ser Thr Asn 130 135 140 Val Lys Ser Ser Ile Ile Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Glu Asn Ser Ser Tyr Pro Val Arg Lys Leu Met Asp Ser 165 170 175 Gly Gly Val Tyr Asp Pro Ser Asn Asp Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly Tyr Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Lys Glu Thr Ile Lys Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Arg Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ile Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Asn Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val 420 425 430 Ile Pro Arg Lys 435 <210> 60 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F-F6 subtype <400> 60 Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Lys Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asp Glu Phe 115 120 125 Ser Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Leu Ser Thr Asn 130 135 140 Val Glu Ser Ser Met Leu Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Glu Ser Cys Cys Tyr Pro Val Arg Lys Leu Ile Asp Pro 165 170 175 Asp Val Val Tyr Asp Pro Ser Asn Tyr Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Glu Glu Thr Ile Glu Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ser Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Ser Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val 420 425 430 Ile Pro Arg Lys 435 <210> 61 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 61 Met Pro Val Val Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Glu Thr Ile Leu Tyr Met Gln Lys Pro Tyr Glu Glu Arg Ser Arg Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Pro Asn Val Trp Ile Met Pro Glu 35 40 45 Arg Asp Thr Ile Gly Thr Lys Pro Asp Glu Phe Gln Val Pro Asp Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Met Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Asn Pro Thr Gly Lys Val Leu Leu Glu Glu Val Ser 100 105 110 Asn Ala Arg Pro Tyr Leu Gly Asp Asp Asp Thr Leu Ile Asn Glu Phe 115 120 125 Phe Pro Val Asn Val Thr Thr Ser Val Asn Ile Lys Phe Ser Thr Asp 130 135 140 Val Glu Ser Ser Ile Ile Ser Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Lys Ala Tyr Cys Thr Pro Leu Val Arg Phe Asn Lys Ser 165 170 175 Asp Lys Leu Ile Glu Pro Ser Asn His Gly Phe Gly Ser Ile Asn Ile 180 185 190 Leu Thr Phe Ser Pro Glu Tyr Glu His Ile Phe Asn Asp Ile Ser Gly 195 200 205 Gly Asn His Asn Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Lys 225 230 235 240 Ala Val Thr His Lys Glu Ser Leu Val Ala Glu Arg Gly Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Glu Asp Leu Asn Ile Ile Pro Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asp Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Arg Glu Val 290 295 300 Asn Thr Ala Pro Pro Gly Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Arg Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Arg Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ile Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Val Lys Val Pro Asp Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Asn Ile Asn Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Ile Lys Phe Cys Lys Ser Ile 420 425 430 Ile Pro Arg Lys 435 <210> 62 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F-F6 subtype <400> 62 Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Lys Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Lys Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asp Glu Phe 115 120 125 Ser Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Leu Ser Thr Asn 130 135 140 Val Glu Ser Ser Met Leu Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Glu Ser Cys Cys Tyr Pro Val Arg Lys Leu Ile Asp Pro 165 170 175 Asp Val Val Tyr Asp Pro Ser Asn Tyr Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Glu Glu Thr Ile Glu Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ser Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Ser Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val 420 425 430 Ile Pro Arg Lys 435 <210> 63 <211> 447 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F-F9 subtype <400> 63 Met Asp Ile Glu Arg Ser Ile Gly Gly Ile Phe Met Pro Phe Glu Ile 1 5 10 15 Asn Ser Phe Asn Tyr Asp Asp Pro Val Asn Asp Glu Thr Ile Leu Tyr 20 25 30 Met Gln Lys Pro Tyr Glu Gly Lys Ser Lys Lys Tyr Tyr Lys Ala Phe 35 40 45 Glu Ile Met Pro Asn Val Trp Ile Met Pro Glu Arg Asp Thr Ile Gly 50 55 60 Thr Glu Pro Ser Asp Phe Gln Val Pro Val Ser Leu Lys Asn Gly Ser 65 70 75 80 Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr Asp Ala Glu Lys Asp 85 90 95 Arg Tyr Leu Lys Ile Met Ile Lys Leu Phe Asn Arg Ile Asn Ser Asn 100 105 110 Pro Ala Gly Glu Val Leu Leu Arg Glu Ile Ser Lys Ala Arg Pro Tyr 115 120 125 Leu Gly Asp Glu His Thr Pro Ile Asn Glu Phe Phe Pro Val Asn Val 130 135 140 Thr Thr Ser Val Asn Ile Lys Phe Ser Asp Asp Val Lys Ser Ser Ile 145 150 155 160 Ile Leu Asn Leu Leu Val Leu Gly Ala Gly Pro Asp Ile Phe Lys Val 165 170 175 Phe Cys Ser Ser Leu Ala Arg Ser Ile Asp Ser Gly Glu Phe Tyr Gln 180 185 190 Pro Ser Asn His Gly Phe Gly Ser Ile Asn Ile Leu Thr Phe Ser Pro 195 200 205 Glu Tyr Glu His Ile Phe Asn Asp Ile Ser Gly Gly Asp His Asn Ser 210 215 220 Ile Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser Leu Ala His Glu Leu 225 230 235 240 Ile His Ala Leu His Gly Leu Tyr Gly Ala Lys Ala Val Thr Tyr Lys 245 250 255 Glu Thr Ile Glu Ala Lys Arg Gly Pro Leu Met Ile Ala Glu Lys Pro 260 265 270 Ile Arg Val Glu Glu Phe Leu Thr Phe Gly Gly Lys Asp Leu Asn Ile 275 280 285 Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn Asn Leu Leu Asp Asn 290 295 300 Tyr Glu Lys Ile Ala Thr Arg Leu Ser Asn Val Asn Thr Ala Pro Ser 305 310 315 320 Gly Tyr Asp Ile Asn Lys Tyr Lys Asp Ile Phe Gln Trp Lys Tyr Gly 325 330 335 Leu Asp Glu Asn Ala Asp Gly Ser Tyr Thr Val Asn Lys Asn Lys Phe 340 345 350 Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr Glu Ile Asp Leu Ala 355 360 365 Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr Phe Ile Glu Tyr Gly 370 375 380 Phe Val Lys Val Pro Asp Leu Leu Asp Asp Asp Ile Tyr Thr Val Ser 385 390 395 400 Glu Gly Phe Asn Ile Asp Asn Leu Ala Val Asn Asn Arg Gly Gln Asn 405 410 415 Ile Asn Leu Asn Pro Lys Ile Ile Gly Ser Ile Pro Asp Glu Gly Leu 420 425 430 Val Glu Lys Ile Val Lys Phe Cys Lys Asn Ile Leu Pro Arg Lys 435 440 445 <210> 64 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 64 Met Pro Val Val Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Glu Thr Ile Leu Tyr Met Gln Lys Pro Tyr Glu Glu Arg Ser Arg Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Pro Asn Val Trp Ile Met Pro Glu 35 40 45 Arg Asp Thr Ile Gly Thr Lys Pro Asp Asp Phe Gln Val Pro Asp Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Met Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Asn Pro Thr Gly Lys Val Leu Leu Glu Glu Val Ser 100 105 110 Asn Ala Arg Pro Tyr Leu Gly Asp Asp Asp Thr Leu Ile Asn Glu Phe 115 120 125 Phe Pro Val Asn Val Thr Thr Ser Val Asn Ile Lys Phe Ser Thr Asp 130 135 140 Val Glu Ser Ser Ile Ile Ser Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Lys Ala Tyr Cys Thr Pro Leu Val Arg Phe Asn Lys Ser 165 170 175 Asp Lys Leu Ile Glu Pro Ser Asn His Gly Phe Gly Ser Ile Asn Ile 180 185 190 Leu Thr Phe Ser Pro Glu Tyr Glu His Ile Phe Asn Asp Ile Ser Gly 195 200 205 Gly Asp His Asn Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Lys 225 230 235 240 Ala Val Thr His Lys Glu Thr Ile Glu Val Lys Arg Gly Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Glu Asp Leu Asn Ile Ile Pro Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asp Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Arg Glu Val 290 295 300 Asn Thr Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Arg Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Arg Asn Lys Phe Asn Gly Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ile Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Val Lys Val Pro Asp Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Asn Ile Asn Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Ile Lys Phe Cys Lys Ser Ile 420 425 430 Ile Pro Arg Lys 435 <210> 65 <211> 428 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F-F7 subtype <400> 65 Met Pro Val Asn Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asn Asn 1 5 10 15 Thr Thr Ile Leu Tyr Met Lys Met Pro Tyr Tyr Glu Asp Ser Asn Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Asp Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Ile Ile Gly Lys Lys Pro Ser Asp Phe Tyr Pro Pro Ile Ser 50 55 60 Leu Asp Ser Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Phe Leu Lys Thr Val Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Gln Val Leu Leu Glu Glu Ile Lys 100 105 110 Asn Gly Lys Pro Tyr Leu Gly Asn Asp His Thr Ala Val Asn Glu Phe 115 120 125 Cys Ala Asn Asn Arg Ser Thr Ser Val Glu Ile Lys Glu Ser Asn Gly 130 135 140 Thr Thr Asp Ser Met Leu Leu Asn Leu Val Ile Leu Gly Pro Gly Pro 145 150 155 160 Asn Ile Leu Glu Cys Ser Thr Phe Pro Val Arg Ile Phe Pro Asn Asn 165 170 175 Ile Ala Tyr Asp Pro Ser Glu Lys Gly Phe Gly Ser Ile Gln Leu Met 180 185 190 Ser Phe Ser Thr Glu Tyr Glu Tyr Ala Phe Asn Asp Asn Thr Asp Leu 195 200 205 Phe Ile Ala Asp Pro Ala Ile Ser Leu Ala His Glu Leu Ile His Val 210 215 220 Leu His Gly Leu Tyr Gly Ala Lys Gly Val Thr Asn Lys Lys Val Ile 225 230 235 240 Glu Val Asp Gln Gly Ala Leu Met Ala Ala Glu Lys Asp Ile Lys Ile 245 250 255 Glu Glu Phe Ile Thr Phe Gly Gly Gln Asp Leu Asn Ile Ile Thr Asn 260 265 270 Ser Thr Asn Gln Lys Ile Tyr Asp Asn Leu Leu Ser Asn Tyr Thr Ala 275 280 285 Ile Ala Ser Arg Leu Ser Gln Val Asn Ile Asn Asn Ser Ala Leu Asn 290 295 300 Thr Thr Tyr Tyr Lys Asn Phe Phe Gln Trp Lys Tyr Gly Leu Asp Gln 305 310 315 320 Asp Ser Asn Gly Asn Tyr Thr Val Asn Ile Ser Lys Phe Asn Ala Ile 325 330 335 Tyr Lys Lys Leu Phe Ser Phe Thr Glu Cys Asp Leu Ala Gln Lys Phe 340 345 350 Gln Val Lys Asn Arg Ser Asn Tyr Leu Phe His Phe Lys Pro Phe Lys 355 360 365 Leu Leu Asp Leu Leu Asp Asp Asn Ile Tyr Ser Ile Ser Glu Gly Phe 370 375 380 Asn Ile Gly Ser Leu Arg Val Asn Asn Asn Gly Gln Asn Ile Asn Leu 385 390 395 400 Asn Ser Arg Ile Val Gly Pro Ile Pro Asp Asn Gly Leu Val Glu Arg 405 410 415 Phe Val Gly Leu Cys Lys Ser Ile Val Ser Lys Lys 420 425 <210> 66 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 66 Met Pro Val Val Ile Asn Ser Phe Asn Tyr Asp Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Met Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Glu Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asn Glu Phe 115 120 125 His Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Ser Ser Thr Asn 130 135 140 Val Glu Ser Ser Ile Ile Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asn Ile Phe Glu Asn Ser Ser Tyr Pro Val Arg Lys Leu Met Asn Ser 165 170 175 Gly Glu Val Tyr Asp Pro Ser Asn Asp Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Lys Glu Thr Ile Lys Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asp Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asn Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ile Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Asn Ile Asn Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Leu Cys Lys Ser Ile 420 425 430 Ile Pro Arg Lys 435 <210> 67 <211> 428 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 67 Met Pro Val Asn Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asn Asn 1 5 10 15 Thr Thr Ile Leu Tyr Met Lys Met Pro Tyr Tyr Glu Asp Ser Asn Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Asp Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Ile Ile Gly Lys Lys Pro Ser Asp Phe Tyr Pro Pro Ile Ser 50 55 60 Leu Asp Ser Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Phe Leu Lys Thr Val Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Gln Val Leu Leu Glu Glu Ile Lys 100 105 110 Asn Gly Lys Pro Tyr Leu Gly Asn Asp His Thr Ala Val Asn Glu Phe 115 120 125 Cys Ala Asn Asn Arg Ser Thr Ser Val Glu Ile Lys Glu Ser Lys Gly 130 135 140 Thr Thr Asp Ser Met Leu Leu Asn Leu Val Ile Leu Gly Pro Gly Pro 145 150 155 160 Asn Ile Leu Glu Cys Ser Thr Phe Pro Val Arg Ile Phe Pro Asn Asn 165 170 175 Ile Ala Tyr Asp Pro Ser Glu Lys Gly Phe Gly Ser Ile Gln Leu Met 180 185 190 Ser Phe Ser Thr Glu Tyr Glu Tyr Ala Phe Asn Asp Asn Thr Asp Leu 195 200 205 Phe Ile Ala Asp Pro Ala Ile Ser Leu Ala His Glu Leu Ile His Val 210 215 220 Leu His Gly Leu Tyr Gly Ala Lys Gly Val Thr Asn Lys Lys Val Ile 225 230 235 240 Glu Val Asp Gln Gly Ala Leu Met Ala Ala Glu Lys Asp Ile Lys Ile 245 250 255 Glu Glu Phe Ile Thr Phe Gly Gly Gln Asp Leu Asn Ile Ile Thr Asn 260 265 270 Ser Thr Asn Gln Lys Ile Tyr Asp Asn Leu Leu Ser Asn Tyr Thr Ala 275 280 285 Ile Ala Ser Arg Leu Ser Gln Val Asn Ile Asn Asn Ser Ala Leu Asn 290 295 300 Thr Thr Tyr Tyr Lys Asn Phe Phe Gln Trp Lys Tyr Gly Leu Asp Gln 305 310 315 320 Asp Ser Asn Gly Asn Tyr Thr Val Asn Ile Ser Lys Phe Asn Ala Ile 325 330 335 Tyr Lys Lys Leu Phe Ser Phe Thr Glu Cys Asp Leu Ala Gln Lys Phe 340 345 350 Gln Val Lys Asn Arg Ser Asn Tyr Leu Phe His Phe Lys Pro Phe Arg 355 360 365 Leu Leu Asp Leu Leu Asp Asp Asn Ile Tyr Ser Ile Ser Glu Gly Phe 370 375 380 Asn Ile Gly Ser Leu Arg Val Asn Asn Asn Gly Gln Asn Ile Asn Leu 385 390 395 400 Asn Ser Arg Ile Val Gly Pro Ile Pro Asp Asn Gly Leu Val Glu Arg 405 410 415 Phe Val Gly Leu Cys Lys Ser Ile Val Ser Lys Lys 420 425 <210> 68 <211> 428 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 68 Met Pro Val Asn Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asn Asn 1 5 10 15 Thr Thr Ile Leu Tyr Met Lys Met Pro Tyr Tyr Glu Asp Ser Asn Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Asp Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Ile Ile Gly Lys Lys Pro Ser Asp Phe Tyr Pro Pro Ile Ser 50 55 60 Leu Asp Ser Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Phe Leu Lys Thr Val Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Gln Val Leu Leu Glu Glu Ile Lys 100 105 110 Asn Gly Lys Pro Tyr Leu Gly Asn Asp His Thr Ala Val Asn Glu Phe 115 120 125 Cys Ala Asn Asn Arg Ser Thr Ser Val Glu Ile Lys Glu Ser Asn Gly 130 135 140 Thr Thr Asp Ser Met Leu Leu Asn Leu Val Ile Leu Gly Pro Gly Pro 145 150 155 160 Asn Ile Leu Glu Cys Ser Thr Phe Pro Val Arg Ile Phe Pro Asn Asn 165 170 175 Ile Ala Tyr Asp Pro Ser Glu Lys Gly Phe Gly Ser Ile Gln Leu Met 180 185 190 Ser Phe Ser Thr Glu Tyr Glu Tyr Ala Phe Asn Asp Asn Thr Asp Leu 195 200 205 Phe Ile Ala Asp Pro Ala Ile Ser Leu Ala His Glu Leu Ile His Val 210 215 220 Leu His Gly Leu Tyr Gly Ala Lys Gly Val Thr Asn Lys Lys Val Ile 225 230 235 240 Glu Val Asp Gln Gly Ala Leu Met Ala Ala Glu Lys Asp Ile Lys Ile 245 250 255 Glu Glu Phe Ile Thr Phe Gly Gly Gln Asp Leu Asn Ile Val Thr Asn 260 265 270 Ser Thr Asn Gln Lys Ile Tyr Asp Asn Leu Leu Ser Asn Tyr Thr Ala 275 280 285 Ile Ala Ser Arg Leu Ser Gln Val Asn Ile Asn Asn Ser Ala Leu Asn 290 295 300 Thr Thr Tyr Tyr Lys Asn Phe Phe Gln Trp Lys Tyr Gly Leu Asp Gln 305 310 315 320 Asp Ser Asn Gly Asn Tyr Thr Val Asn Ile Ser Lys Phe Asn Ala Ile 325 330 335 Tyr Lys Lys Leu Phe Ser Phe Thr Glu Cys Asp Leu Ala Gln Lys Phe 340 345 350 Gln Val Lys Asn Arg Ser Asn Tyr Leu Phe His Phe Lys Pro Phe Lys 355 360 365 Leu Leu Asp Leu Leu Asp Asp Asn Ile Tyr Ser Ile Ser Glu Gly Phe 370 375 380 Asn Ile Gly Ser Leu Arg Val Asn Asn Asn Gly Gln Asn Ile Asn Leu 385 390 395 400 Asn Ser Arg Ile Val Gly Pro Ile Pro Asp Asn Gly Leu Val Glu Arg 405 410 415 Phe Val Gly Leu Cys Lys Ser Ile Val Ser Lys Lys 420 425 <210> 69 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 69 Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asp Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Glu Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Glu Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Glu His Thr Pro Ile Asn Glu Phe 115 120 125 His Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Ser Ser Thr Asn 130 135 140 Val Lys Ser Ser Ile Ile Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Glu Asn Ser Ser Tyr Pro Val Arg Lys Leu Met Asp Ser 165 170 175 Gly Gly Val Tyr Asp Pro Ser Asn Asp Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly Tyr Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Lys Glu Thr Ile Lys Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Arg Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ile Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Asn Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val 420 425 430 Ile Pro Arg Lys 435 <210> 70 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/G <400> 70 Met Pro Val Asn Ile Lys Phe Asn Tyr Asn Asp Pro Ile Asn Asn Asp 1 5 10 15 Asp Ile Ile Met Met Glu Pro Phe Asn Asp Pro Gly Pro Gly Thr Tyr 20 25 30 Tyr Lys Ala Phe Arg Ile Ile Asp Arg Ile Trp Ile Val Pro Glu Arg 35 40 45 Phe Thr Tyr Gly Phe Gln Pro Asp Gln Phe Asn Ala Ser Thr Gly Val 50 55 60 Phe Ser Lys Asp Val Tyr Glu Tyr Tyr Asp Pro Thr Tyr Leu Lys Thr 65 70 75 80 Asp Ala Glu Lys Asp Lys Phe Leu Lys Thr Met Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Lys Pro Ser Gly Gln Arg Leu Leu Asp Met Ile Val 100 105 110 Asp Ala Ile Pro Tyr Leu Gly Asn Ala Ser Thr Pro Pro Asp Lys Phe 115 120 125 Ala Ala Asn Val Ala Asn Val Ser Ile Asn Lys Lys Ile Ile Gln Pro 130 135 140 Gly Ala Glu Asp Gln Ile Lys Gly Leu Met Thr Asn Leu Ile Ile Phe 145 150 155 160 Gly Pro Gly Pro Val Leu Ser Asp Asn Phe Thr Asp Ser Met Ile Met 165 170 175 Asn Gly His Ser Pro Ile Ser Glu Gly Phe Gly Ala Arg Met Met Ile 180 185 190 Arg Phe Cys Pro Ser Cys Leu Asn Val Phe Asn Asn Val Gln Glu Asn 195 200 205 Lys Asp Thr Ser Ile Phe Ser Arg Arg Ala Tyr Phe Ala Asp Pro Ala 210 215 220 Leu Thr Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr Gly 225 230 235 240 Ile Lys Ile Ser Asn Leu Pro Ile Thr Pro Asn Thr Lys Glu Phe Phe 245 250 255 Met Gln His Ser Asp Pro Val Gln Ala Glu Glu Leu Tyr Thr Phe Gly 260 265 270 Gly His Asp Pro Ser Val Ile Ser Pro Ser Thr Asp Met Asn Ile Tyr 275 280 285 Asn Lys Ala Leu Gln Asn Phe Gln Asp Ile Ala Asn Arg Leu Asn Ile 290 295 300 Val Ser Ser Ala Gln Gly Ser Gly Ile Asp Ile Ser Leu Tyr Lys Gln 305 310 315 320 Ile Tyr Lys Asn Lys Tyr Asp Phe Val Glu Asp Pro Asn Gly Lys Tyr 325 330 335 Ser Val Asp Lys Asp Lys Phe Asp Lys Leu Tyr Lys Ala Leu Met Phe 340 345 350 Gly Phe Thr Glu Thr Asn Leu Ala Gly Glu Tyr Gly Ile Lys Thr Arg 355 360 365 Tyr Ser Tyr Phe Ser Glu Tyr Leu Pro Pro Ile Lys Thr Glu Lys Leu 370 375 380 Leu Asp Asn Thr Ile Tyr Thr Gln Asn Glu Gly Phe Asn Ile Ala Ser 385 390 395 400 Lys Asn Leu Lys Thr Glu Phe Asn Gly Gln Asn Lys Ala Val Asn Lys 405 410 415 Glu Ala Tyr Glu Glu Ile Ser Leu Glu His Leu Val Ile Tyr Arg Ile 420 425 430 Ala Met Cys Lys Pro Val Met Tyr Lys 435 440 <210> 71 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> EGFP <400> 71 Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 1 5 10 15 Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly 20 25 30 Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile 35 40 45 Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 50 55 60 Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys 65 70 75 80 Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu 85 90 95 Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu 100 105 110 Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly 115 120 125 Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 130 135 140 Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn 145 150 155 160 Gly Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser 165 170 175 Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly 180 185 190 Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu 195 200 205 Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe 210 215 220 Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys 225 230 235 <210> 72 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> EYFP <400> 72 Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 1 5 10 15 Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly 20 25 30 Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile 35 40 45 Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 50 55 60 Phe Gly Tyr Gly Leu Gln Cys Phe Ala Arg Tyr Pro Asp His Met Lys 65 70 75 80 Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu 85 90 95 Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu 100 105 110 Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly 115 120 125 Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 130 135 140 Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn 145 150 155 160 Gly Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser 165 170 175 Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly 180 185 190 Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Tyr Gln Ser Ala Leu 195 200 205 Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe 210 215 220 Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys 225 230 235 <210> 73 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> ECFP <400> 73 Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 1 5 10 15 Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly 20 25 30 Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile 35 40 45 Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 50 55 60 Leu Thr Trp Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys 65 70 75 80 Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu 85 90 95 Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu 100 105 110 Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly 115 120 125 Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 130 135 140 Asn Tyr Ile Ser His Asn Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn 145 150 155 160 Gly Ile Lys Ala Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser 165 170 175 Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly 180 185 190 Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu 195 200 205 Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe 210 215 220 Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys 225 230 235 <210> 74 <211> 236 <212> PRT <213> Artificial Sequence <220> <223> ERFP <400> 74 Met Ser Cys Ser Lys Asn Val Ile Lys Glu Phe Met Arg Phe Lys Val 1 5 10 15 Arg Met Glu Gly Thr Val Asn Gly His Glu Phe Glu Ile Glu Gly Glu 20 25 30 Gly Glu Gly Arg Pro Tyr Glu Gly His Asn Thr Val Lys Leu Lys Val 35 40 45 Thr Lys Gly Gly Pro Leu Pro Phe Ala Trp Asp Ile Leu Ser Pro Gln 50 55 60 Phe Gln Tyr Gly Ser Lys Val Tyr Val Lys His Pro Ala Asp Ile Pro 65 70 75 80 Asp Tyr Lys Lys Leu Ser Phe Pro Glu Gly Phe Lys Trp Glu Arg Val 85 90 95 Met Asn Phe Glu Asp Gly Gly Val Val Thr Val Thr Gln Asp Ser Ser 100 105 110 Leu Gln Asp Gly Cys Phe Ile Tyr Lys Val Lys Phe Ile Gly Val Asn 115 120 125 Phe Pro Ser Asp Gly Pro Val Met Gln Lys Lys Thr Met Gly Trp Glu 130 135 140 Ala Ser Thr Glu Arg Leu Tyr Pro Arg Asp Gly Val Leu Lys Gly Glu 145 150 155 160 Ile His Lys Ala Leu Lys Leu Lys Asp Gly Gly His Tyr Leu Val Glu 165 170 175 Phe Lys Thr Ile Tyr Met Ala Lys Lys Pro Val Gln Leu Pro Gly Tyr 180 185 190 Tyr Tyr Val Asp Ser Lys Leu Asp Ile Thr Ser His Asn Lys Asp Tyr 195 200 205 Thr Ile Val Glu Gln Tyr Glu Arg Thr Glu Gly Arg His His Leu Phe 210 215 220 Leu Lys Ala Glu Leu Gly Ser Asn Val Gly Glu Arg 225 230 235 <210> 75 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> EBFP <400> 75 Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 1 5 10 15 Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly 20 25 30 Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile 35 40 45 Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 50 55 60 Leu Thr His Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys 65 70 75 80 Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu 85 90 95 Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu 100 105 110 Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly 115 120 125 Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 130 135 140 Asn Phe Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn 145 150 155 160 Gly Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser 165 170 175 Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly 180 185 190 Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu 195 200 205 Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe 210 215 220 Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys 225 230 235 <210> 76 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> His10 <400> 76 His His His His His His His His His His 1 5 10 <210> 77 <211> 26 <212> PRT <213> Artificial Sequence <220> <223> LAH4 <400> 77 Lys Lys Ala Leu Leu Ala Leu Ala Leu His His Leu Ala His Leu Ala 1 5 10 15 Leu His Leu Ala Leu Ala Leu Lys Lys Ala 20 25 <210> 78 <211> 26 <212> PRT <213> Artificial Sequence <220> <223> LAH4-L1 <400> 78 Lys Lys Ala Leu Leu Ala His Ala Leu His Leu Leu Ala Leu Leu Ala 1 5 10 15 Leu His Leu Ala His Ala Leu Lys Lys Ala 20 25 <210> 79 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> K6p <400> 79 Cys His Lys Lys Lys Lys Lys Lys His Cys 1 5 10 <210> 80 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> H5WYG <400> 80 Gly Leu Phe His Ala Ile Ala His Phe Ile His Gly Gly Trp His Gly 1 5 10 15 Leu Ile His Gly Trp Tyr Gly 20 <210> 81 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> diINF-7 <400> 81 Gly Leu Phe His Ala Ile Ala His Phe Ile His Gly Gly Trp His Gly 1 5 10 15 Leu Ile His Gly Trp Tyr Gly 20 <210> 82 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> TAT <400> 82 Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Arg Arg Gln 1 5 10 <210> 83 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Glycoprotein <400> 83 Leu Ala Ser Thr Leu Thr Arg Trp Ala His Tyr Asn Ala Leu Ile Arg 1 5 10 15 Ala Phe <210> 84 <211> 26 <212> PRT <213> Artificial Sequence <220> <223> Melittin <400> 84 Gly Ile Gly Ala Val Leu Lys Val Leu Thr Thr Gly Leu Pro Ala Leu 1 5 10 15 Ile Ser Trp Ile Lys Arg Lys Arg Gln Gln 20 25 <210> 85 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> MT20 <400> 85 Gly Ile Gly Ala Val Leu Lys Val Leu Thr Thr Gly Leu Pro Ala Leu 1 5 10 15 Ile Ser Trp Ile 20 <210> 86 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Penetratin <400> 86 Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys 1 5 10 15 <210> 87 <211> 22 <212> PRT <213> Artificial Sequence <220> <223> R6-Penetratin <400> 87 Arg Arg Arg Arg Arg Arg Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg 1 5 10 15 Arg Met Lys Trp Lys Lys 20 <210> 88 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Transportan <400> 88 Gly Trp Thr Leu Asn Ser Ala Gly Tyr Leu Leu Gly Lys Ile Asn Leu 1 5 10 15 Lys Ala Leu Ala Ala Leu Ala Lys Lys Ile Leu 20 25 <210> 89 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> TP10 <400> 89 Ala Gly Tyr Leu Leu Gly Lys Ile Asn Leu Lys Ala Leu Ala Ala Leu 1 5 10 15 Ala Lys Lys Ile Leu 20 <210> 90 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> EB1 <400> 90 Leu Ile Arg Leu Trp Ser His Leu Ile His Ile Trp Phe Gln Asn Arg 1 5 10 15 Arg Leu Lys Trp Lys Lys Lys 20 <210> 91 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Bovine prion protein <400> 91 Met Val Lys Ser Lys Ile Gly Ser Trp Ile Leu Val Leu Phe Val Ala 1 5 10 15 Met Trp Ser Asp Val Gly Leu Cys Lys Lys Arg Pro Lys Pro 20 25 30 <210> 92 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Aurein 1.2 <400> 92 Gly Leu Phe Asp Ile Ile Lys Lys Ile Ala Glu Ser Phe 1 5 10 <210> 93 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Proline-rich protein <400> 93 Cys Phe Val Arg Leu Pro Pro Pro Val Arg Leu Pro Pro Pro Val Arg 1 5 10 15 Leu Pro Pro Pro 20 <210> 94 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> GALA <400> 94 Trp Glu Ala Ala Leu Ala Glu Ala Leu Ala Glu Ala Leu Ala Glu His 1 5 10 15 Leu Ala Glu Ala Leu Ala Glu Ala Leu Glu Ala Leu Ala Ala 20 25 30 <210> 95 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> KALA <400> 95 Trp Glu Ala Lys Leu Ala Lys Ala Leu Ala Lys Ala Leu Ala Lys His 1 5 10 15 Leu Ala Lys Ala Leu Ala Lys Ala Leu Lys Ala Cys Glu Ala 20 25 30 <210> 96 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> JTS-1 <400> 96 Gly Leu Phe Glu Ala Leu Leu Glu Leu Leu Glu Ser Leu Trp Glu Leu 1 5 10 15 Leu Leu Glu Ala 20 <210> 97 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> ppTG1 <400> 97 Gly Leu Phe Lys Ala Leu Leu Lys Leu Leu Lys Ser Leu Trp Lys Leu 1 5 10 15 Leu Leu Lys Ala 20 <210> 98 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> ppTG20 <400> 98 Gly Leu Arg Lys Ala Leu Leu Arg Leu Leu Arg Ser Leu Trp Arg Leu 1 5 10 15 Leu Leu Arg Ala 20 <210> 99 <211> 427 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype RBD <400> 99 Met Lys Asn Ile Ile Asn Thr Ser Ile Leu Asn Leu Arg Tyr Glu Ser 1 5 10 15 Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile Gly 20 25 30 Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu Phe 35 40 45 Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile Val 50 55 60 Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg Ile 65 70 75 80 Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile Ile 85 90 95 Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr Gly 100 105 110 Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg Val 115 120 125 Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn Arg 130 135 140 Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys Ile 145 150 155 160 Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu Gly 165 170 175 Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys Arg 180 185 190 Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp Lys 195 200 205 Glu Leu Asn Glu Lys Glu Ile Lys Asp Leu Tyr Asp Asn Gln Ser Asn 210 215 220 Ser Gly Ile Leu Lys Asp Phe Trp Gly Asp Tyr Leu Gln Tyr Asp Lys 225 230 235 240 Pro Tyr Tyr Met Leu Asn Leu Tyr Asp Pro Asn Lys Tyr Val Asp Val 245 250 255 Asn Asn Val Gly Ile Arg Gly Tyr Met Tyr Leu Lys Gly Pro Arg Gly 260 265 270 Ser Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser Leu Tyr Arg Gly 275 280 285 Thr Lys Phe Ile Ile Lys Lys Tyr Ala Ser Gly Asn Lys Asp Asn Ile 290 295 300 Val Arg Asn Asn Asp Arg Val Tyr Ile Asn Val Val Val Lys Asn Lys 305 310 315 320 Glu Tyr Arg Leu Ala Thr Asn Ala Ser Gln Ala Gly Val Glu Lys Ile 325 330 335 Leu Ser Ala Leu Glu Ile Pro Asp Val Gly Asn Leu Ser Gln Val Val 340 345 350 Val Met Lys Ser Lys Asn Asp Gln Gly Ile Thr Asn Lys Cys Lys Met 355 360 365 Asn Leu Gln Asp Asn Asn Gly Asn Asp Ile Gly Phe Ile Gly Phe His 370 375 380 Gln Phe Asn Asn Ile Ala Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg 385 390 395 400 Gln Ile Glu Arg Ser Ser Arg Thr Leu Gly Cys Ser Trp Glu Phe Ile 405 410 415 Pro Val Asp Asp Gly Trp Gly Glu Arg Pro Leu 420 425 <210> 100 <211> 427 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype RBD <400> 100 Met Lys Asn Ile Val Asn Thr Ser Ile Leu Ser Ile Val Tyr Lys Lys 1 5 10 15 Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile Gly 20 25 30 Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu Ile 35 40 45 Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile Val 50 55 60 Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys Ile 65 70 75 80 Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile Ile 85 90 95 Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr Gly 100 105 110 Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg Val 115 120 125 Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn Arg 130 135 140 Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys Ile 145 150 155 160 Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu Gly 165 170 175 Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys Arg 180 185 190 Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp Lys 195 200 205 Glu Leu Asn Glu Lys Glu Ile Lys Asp Leu Tyr Asp Ser Gln Ser Asn 210 215 220 Ser Gly Ile Leu Lys Asp Phe Trp Gly Asn Tyr Leu Gln Tyr Asp Lys 225 230 235 240 Pro Tyr Tyr Met Leu Asn Leu Phe Asp Pro Asn Lys Tyr Val Asp Val 245 250 255 Asn Asn Ile Gly Ile Arg Gly Tyr Met Tyr Leu Lys Gly Pro Arg Gly 260 265 270 Ser Val Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr Leu Tyr Glu Gly 275 280 285 Thr Lys Phe Ile Ile Lys Lys Tyr Ala Ser Gly Asn Glu Asp Asn Ile 290 295 300 Val Arg Asn Asn Asp Arg Val Tyr Ile Asn Val Val Val Lys Asn Lys 305 310 315 320 Glu Tyr Arg Leu Ala Thr Asn Ala Ser Gln Ala Gly Val Glu Lys Ile 325 330 335 Leu Ser Ala Leu Glu Ile Pro Asp Val Gly Asn Leu Ser Gln Val Val 340 345 350 Val Met Lys Ser Lys Asp Asp Gln Gly Ile Arg Asn Lys Cys Lys Met 355 360 365 Asn Leu Gln Asp Asn Asn Gly Asn Asp Ile Gly Phe Ile Gly Phe His 370 375 380 Leu Tyr Asp Asn Ile Ala Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg 385 390 395 400 Gln Val Gly Lys Ala Ser Arg Thr Phe Gly Cys Ser Trp Glu Phe Ile 405 410 415 Pro Val Asp Asp Gly Trp Gly Glu Ser Ser Leu 420 425 <210> 101 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 101 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Arg Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 102 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype RBD <400> 102 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile 1 5 10 15 Gly Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Glu Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Leu Tyr Asp Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 103 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype RBD <400> 103 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Arg Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 104 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A5 subtype RBD <400> 104 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Glu Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Lys Ile Glu Ile Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Ala Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Ile Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Ile Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Val Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Asp Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 105 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A3 subtype RBD <400> 105 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile 1 5 10 15 Gly Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Met Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Pro Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Val Gly Phe His Leu Tyr Asp Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 106 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 106 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile 1 5 10 15 Gly Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Arg Val Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Glu Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Leu Tyr Asp Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 107 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 107 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile 1 5 10 15 Gly Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Glu Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Leu Tyr Asp Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 108 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 108 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile 1 5 10 15 Gly Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Glu Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Leu Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 109 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype RBD <400> 109 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Ile Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Asn Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 110 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype RBD <400> 110 Met Asn Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn 1 5 10 15 Ile Gly Ser Arg Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln 20 25 30 Leu Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala 35 40 45 Ile Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile 50 55 60 Lys Ile Pro Lys Tyr Phe Ser Glu Ile Ser Leu Asn Asn Glu Tyr Thr 65 70 75 80 Ile Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn 85 90 95 Tyr Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln 100 105 110 Arg Val Val Phe Lys Tyr Ser Gln Met Val Ala Ile Ser Asp Tyr Ile 115 120 125 Asn Arg Trp Ile Phe Ile Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser 130 135 140 Lys Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn 145 150 155 160 Leu Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly 165 170 175 Cys Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe 180 185 190 Asp Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp 195 200 205 Phe Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn 210 215 220 Leu Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg 225 230 235 240 Gly Tyr Met Tyr Leu Lys Gly Ser Arg Ser Thr Leu Leu Thr Thr Asn 245 250 255 Ile Tyr Leu Asn Ser Gly Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys 260 265 270 Lys Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg 275 280 285 Val Tyr Ile Asn Val Val Val Asn Asn Lys Glu Tyr Arg Leu Ala Thr 290 295 300 Asn Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile 305 310 315 320 Pro Asp Ile Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn 325 330 335 Asp Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn 340 345 350 Gly Asn Asp Ile Gly Phe Ile Gly Phe His Lys Phe Asn Asp Ile Tyr 355 360 365 Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Ile Ser Ser 370 375 380 Arg Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp 385 390 395 400 Gly Glu <210> 111 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 111 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Glu Ile Tyr Asn 1 5 10 15 Gly Asp Lys Val Tyr Tyr Asn Ser Ile Asp Lys Asn Gln Ile Arg Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Lys Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Phe Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Ile Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro His Arg Tyr Ile Val Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Ser Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Asp Asn Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Arg 385 390 395 400 Glu <210> 112 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype RBD <400> 112 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Arg Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 113 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 113 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Ile Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Asn Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 114 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype RBD <400> 114 Met Asp Glu Leu Ile Asp Leu Ser Arg Tyr Gly Ala Glu Ile Tyr Arg 1 5 10 15 Gly Asp Lys Val Phe Tyr Asn Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Leu Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Gly Arg Tyr Ile Val Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Leu Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Leu Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 115 <211> 425 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 115 Met Ile Ile Asn Thr Ser Ile Leu Asn Leu Arg Tyr Glu Ser Asn His 1 5 10 15 Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile Gly Ser Lys 20 25 30 Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu Phe Asn Leu 35 40 45 Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile Val Tyr Asn 50 55 60 Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg Ile Pro Lys 65 70 75 80 Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile Ile Asn Cys 85 90 95 Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr Gly Glu Ile 100 105 110 Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg Val Val Phe 115 120 125 Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn Arg Trp Ile 130 135 140 Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys Ile Tyr Ile 145 150 155 160 Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu Gly Asn Ile 165 170 175 His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys Arg Asp Thr 180 185 190 His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp Lys Glu Leu 195 200 205 Asn Glu Lys Glu Ile Lys Asp Leu Tyr Asp Asn Gln Ser Asn Ser Gly 210 215 220 Ile Leu Lys Asp Phe Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr 225 230 235 240 Tyr Met Leu Asn Leu Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn 245 250 255 Val Gly Ile Arg Gly Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val 260 265 270 Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser Leu Tyr Arg Gly Thr Lys 275 280 285 Phe Ile Ile Lys Lys Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg 290 295 300 Asn Asn Asp Arg Val Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr 305 310 315 320 Arg Leu Ala Thr Asn Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser 325 330 335 Ala Leu Glu Ile Pro Asp Val Gly Asn Leu Ser Gln Val Val Val Met 340 345 350 Lys Ser Lys Asn Asp Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu 355 360 365 Gln Asp Asn Asn Gly Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe 370 375 380 Asn Asn Ile Ala Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile 385 390 395 400 Glu Arg Ser Ser Arg Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val 405 410 415 Asp Asp Gly Trp Gly Glu Arg Pro Leu 420 425 <210> 116 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A1-A2 <400> 116 Arg Gly Ser Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser 1 5 10 <210> 117 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A2-A2 <400> 117 Arg Gly Ser Val Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr 1 5 10 <210> 118 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A3-A2 <400> 118 Arg Gly Ser Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Thr 1 5 10 <210> 119 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A4-A2 <400> 119 Arg Gly Asp Asn Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser 1 5 10 <210> 120 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A5-A2 <400> 120 Arg Gly Ser Val Ile Val Thr Asn Ile Tyr Leu Asn Ser Ser 1 5 10 <210> 121 <211> 437 <212> PRT <213> Artificial Sequence <220> <223> BoNTs LC/amino acid sequence 1 <400> 121 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe 435 <210> 122 <211> 451 <212> PRT <213> Artificial Sequence <220> <223> BoNTs LC-A2/amino acid sequence 2 <400> 122 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Arg Gly Ser Val Val Thr Thr Asn Ile Tyr Leu 435 440 445 Asn Ser Thr 450 <210> 123 <211> 464 <212> PRT <213> Artificial Sequence <220> <223> BoNTs LC-Aurein 1.2-A2/amino acid sequence 3 <400> 123 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Gly Leu Phe Asp Ile Ile Lys Lys Ile Ala Glu 435 440 445 Ser Phe Arg Gly Ser Val Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr 450 455 460 <210> 124 <211> 461 <212> PRT <213> Artificial Sequence <220> <223> Recombinant botulinum neurotoxin amino acid sequence 4 <400> 124 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe His His His His His His His His His His Arg 435 440 445 Gly Ser Val Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr 450 455 460 <210> 125 <211> 474 <212> PRT <213> Artificial Sequence <220> <223> Aurein 1.2-BoNTs LC-His10-A2/amino acid sequence 5 <400> 125 Met Gly Leu Phe Asp Ile Ile Lys Lys Ile Ala Glu Ser Phe Pro Phe 1 5 10 15 Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly Val Asp Ile 20 25 30 Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro Val Lys Ala 35 40 45 Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg Asp Thr Phe 50 55 60 Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu Ala Lys Gln 65 70 75 80 Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr Asp Asn Glu 85 90 95 Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu Arg Ile Tyr 100 105 110 Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val Arg Gly Ile 115 120 125 Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys Val Ile Asp 130 135 140 Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr Arg Ser Glu 145 150 155 160 Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile Ile Gln Phe 165 170 175 Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr Arg Asn Gly 180 185 190 Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe Thr Phe Gly 195 200 205 Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu Gly Ala Gly 210 215 220 Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu Leu Ile His 225 230 235 240 Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn Arg Val Phe 245 250 255 Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu Glu Val Ser 260 265 270 Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys Phe Ile Asp 275 280 285 Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn Lys Phe Lys 290 295 300 Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile Gly Thr Thr 305 310 315 320 Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys Tyr Leu Leu 325 330 335 Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu Lys Phe Asp 340 345 350 Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp Asn Phe Val 355 360 365 Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn Phe Asp Lys 370 375 380 Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr Thr Ile Lys 385 390 395 400 Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn Phe Asn Gly 405 410 415 Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu Lys Asn Phe 420 425 430 Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg Gly Ile Ile 435 440 445 Pro Phe His His His His His His His His His His Arg Gly Ser Val 450 455 460 Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr 465 470 <210> 126 <211> 474 <212> PRT <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/amino acid sequence 6 <400> 126 Met Gly Leu Phe Asp Ile Ile Lys Lys Ile Ala Glu Ser Phe His His 1 5 10 15 His His His His His His His His Pro Phe Val Asn Lys Gln Phe Asn 20 25 30 Tyr Lys Asp Pro Val Asn Gly Val Asp Ile Ala Tyr Ile Lys Ile Pro 35 40 45 Asn Ala Gly Gln Met Gln Pro Val Lys Ala Phe Lys Ile His Asn Lys 50 55 60 Ile Trp Val Ile Pro Glu Arg Asp Thr Phe Thr Asn Pro Glu Glu Gly 65 70 75 80 Asp Leu Asn Pro Pro Pro Glu Ala Lys Gln Val Pro Val Ser Tyr Tyr 85 90 95 Asp Ser Thr Tyr Leu Ser Thr Asp Asn Glu Lys Asp Asn Tyr Leu Lys 100 105 110 Gly Val Thr Lys Leu Phe Glu Arg Ile Tyr Ser Thr Asp Leu Gly Arg 115 120 125 Met Leu Leu Thr Ser Ile Val Arg Gly Ile Pro Phe Trp Gly Gly Ser 130 135 140 Thr Ile Asp Thr Glu Leu Lys Val Ile Asp Thr Asn Cys Ile Asn Val 145 150 155 160 Ile Gln Pro Asp Gly Ser Tyr Arg Ser Glu Glu Leu Asn Leu Val Ile 165 170 175 Ile Gly Pro Ser Ala Asp Ile Ile Gln Phe Glu Cys Lys Ser Phe Gly 180 185 190 His Asp Val Leu Asn Leu Thr Arg Asn Gly Tyr Gly Ser Thr Gln Tyr 195 200 205 Ile Arg Phe Ser Pro Asp Phe Thr Phe Gly Phe Glu Glu Ser Leu Glu 210 215 220 Val Asp Thr Asn Pro Leu Leu Gly Ala Gly Lys Phe Ala Thr Asp Pro 225 230 235 240 Ala Val Thr Leu Ala His Glu Leu Ile His Ala Glu His Arg Leu Tyr 245 250 255 Gly Ile Ala Ile Asn Pro Asn Arg Val Phe Lys Val Asn Thr Asn Ala 260 265 270 Tyr Tyr Glu Met Ser Gly Leu Glu Val Ser Phe Glu Glu Leu Arg Thr 275 280 285 Phe Gly Gly His Asp Ala Lys Phe Ile Asp Ser Leu Gln Glu Asn Glu 290 295 300 Phe Arg Leu Tyr Tyr Tyr Asn Lys Phe Lys Asp Val Ala Ser Thr Leu 305 310 315 320 Asn Lys Ala Lys Ser Ile Ile Gly Thr Thr Ala Ser Leu Gln Tyr Met 325 330 335 Lys Asn Val Phe Lys Glu Lys Tyr Leu Leu Ser Glu Asp Thr Ser Gly 340 345 350 Lys Phe Ser Val Asp Lys Leu Lys Phe Asp Lys Leu Tyr Lys Met Leu 355 360 365 Thr Glu Ile Tyr Thr Glu Asp Asn Phe Val Asn Phe Phe Lys Val Ile 370 375 380 Asn Arg Lys Thr Tyr Leu Asn Phe Asp Lys Ala Val Phe Arg Ile Asn 385 390 395 400 Ile Val Pro Asp Glu Asn Tyr Thr Ile Lys Asp Gly Phe Asn Leu Lys 405 410 415 Gly Ala Asn Leu Ser Thr Asn Phe Asn Gly Gln Asn Thr Glu Ile Asn 420 425 430 Ser Arg Asn Phe Thr Arg Leu Lys Asn Phe Thr Gly Leu Phe Glu Phe 435 440 445 Tyr Lys Leu Leu Cys Val Arg Gly Ile Ile Pro Phe Arg Gly Ser Val 450 455 460 Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr 465 470 <210> 127 <211> 1311 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC/DNA sequence 1 <400> 127 atgcccttcg tcaacaaaca gttcaattat aaggatccgg taaatggggt agacatagca 60 tacataaaga ttcccaacgc aggacaaatg caaccggtga aggcgttcaa aattcataat 120 aagatatggg tcatacctga gcgggacacc tttacaaacc ccgaggaagg tgatcttaac 180 ccgcctccgg aagcgaaaca agttcctgtt tcctactatg actcaactta cctgtcaact 240 gataatgaga aggacaatta tttaaaaggc gtcacgaagt tattcgaaag aatatattca 300 actgacctgg gacgcatgct gctgacttct atcgttagag ggataccatt ctggggaggt 360 tctactattg atactgagct gaaggtaatt gacaccaact gtattaatgt tattcaacct 420 gatggttcct acagatcaga agagttaaat cttgtcatta taggtcccag tgcagacatc 480 attcagtttg aatgtaaatc tttcgggcac gatgtgctta atttgaccag aaatggttat 540 ggaagtactc aatacattag attttcgccg gactttacat tcggctttga ggaaagtctt 600 gaagtggaca cgaacccttt gctgggggca ggtaaatttg ctacggatcc agcggtcacc 660 cttgcacacg agcttatcca tgcggagcac cgcttatacg gcatagcaat aaaccccaat 720 cgtgtcttta aggtaaatac gaacgcttac tatgaaatgt ctggtctgga ggtaagcttt 780 gaagaactgc gtactttcgg ggggcacgac gccaagttca tcgactccct gcaagagaat 840 gagtttcggc tgtactatta caataaattc aaggacgtag cgagcacctt aaataaagcc 900 aaaagcatca ttggcactac tgcctcttta cagtacatga agaacgtatt taaagagaag 960 tatcttttaa gcgaagatac gtcgggcaag tttagcgtcg ataagttaaa gtttgacaag 1020 ttatataaga tgttgacgga gatttataca gaggataact tcgttaactt cttcaaagta 1080 ataaaccgta aaacgtacct gaacttcgat aaggctgtgt tccgtataaa catcgtgccg 1140 gacgagaact acacaattaa ggacggcttt aacttaaaag gggcaaacct gagtaccaac 1200 ttcaacggcc aaaatactga gataaattca cgtaatttta cccgtttgaa gaattttacg 1260 ggcctgttcg agttttacaa gttactttgt gtccgtggca tcattccctt c 1311 <210> 128 <211> 1353 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-A2/DNA sequence 2 <400> 128 atgcctttcg ttaataagca gttcaactat aaggatcctg tcaacggtgt cgacatcgcg 60 tacataaaaa tcccgaatgc tggtcaaatg cagccggtaa aagccttcaa aattcacaat 120 aagatctggg taatacctga gcgtgatacg ttcacgaatc cagaggaagg tgatttgaac 180 ccgcctccag aggcaaaaca ggttcctgta tcgtattacg atagtaccta cctgagtacg 240 gacaacgaga aagacaatta tttaaagggt gtcacgaaac tgtttgagcg gatttactca 300 acggatttgg gaagaatgct gttgacgtca atcgtaagag gtattccttt ctggggtggt 360 tcaaccatag atacagaact gaaggtcatt gatactaatt gtataaatgt catacagcct 420 gatggttctt atcgctccga agaactgaac ttggttatta tagggccatc cgctgatata 480 attcagtttg agtgtaaaag tttcggccac gacgtcttaa acttgacaag aaacgggtac 540 gggtctactc aatatattcg cttcagccct gatttcactt ttggcttcga ggaatcgtta 600 gaagtggaca caaatccgct tctgggggcg ggaaaatttg ctacggaccc cgcggtcacg 660 cttgctcacg agttaattca tgcagagcat cggttatacg gcatcgccat caatccaaac 720 cgggttttca aggtaaatac gaacgcctac tacgaaatga gcggattaga ggtatcgttc 780 gaagaacttc ggacgtttgg cggacatgac gccaaattca tagattcact gcaagaaaat 840 gaatttagat tgtattatta caacaagttt aaagacgtgg cgtccaccct taataaggcc 900 aagtctatca taggtacaac agcatcgtta cagtatatga agaacgtttt caaggaaaaa 960 taccttttgt ctgaagacac cagcgggaag ttctcggttg ataaactgaa gtttgacaaa 1020 ctgtacaaga tgttgacaga aatctacact gaggacaatt ttgtcaattt ttttaaggtc 1080 attaatcgta agacatatct gaatttcgac aaagcggtat tccgtataaa catcgtgcca 1140 gatgagaatt atacaattaa ggacggcttc aacttaaaag gtgctaacct tagcaccaac 1200 tttaacggcc agaataccga gattaattcc cgcaacttca cccgcttaaa aaactttaca 1260 ggactttttg aattttacaa attgctgtgt gtgcggggca taatcccttt tagagggtct 1320 gtggtcacca caaatatata cctgaacagt act 1353 <210> 129 <211> 1392 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-Aurein 1.2-A2/DNA sequence 3 <400> 129 atgcctttcg taaacaaaca attcaactat aaagaccctg tcaatggggt agacatcgcg 60 tacataaaga ttcccaatgc gggccagatg caaccagtca aagcctttaa gatccacaac 120 aagatctggg tgattccaga acgcgacacg ttcaccaatc ctgaagaagg cgacttgaat 180 ccaccgccag aggccaaaca agtaccggta agctattacg actccaccta tttatcgaca 240 gataatgaga aagacaacta cttaaaaggt gtaaccaagt tatttgagcg tatatactcg 300 actgatctgg gaagaatgct gttaacttct attgttagag gaattccttt ttggggtgga 360 tccactatcg acaccgagtt aaaagttatt gacacaaatt gtataaacgt tatccagccc 420 gacggttcct accgttccga agaactgaac cttgtcatca ttggtccctc ggcagatata 480 attcaatttg aatgtaagag tttcgggcac gatgtcttga atctgactag aaacgggtat 540 ggatcaacac agtatattag attttcgccc gacttcacgt tcggattcga ggaaagttta 600 gaagtcgata ctaacccctt gttaggtgcg gggaaatttg caaccgaccc tgctgttacc 660 ttggcccacg agttgatcca cgcagagcat cggctttatg gaattgctat taacccaaac 720 cgcgtcttta aagtaaacac taacgcctac tatgagatga gtggcttgga ggtgagtttt 780 gaagagctta gaacctttgg tggccatgat gctaagttta tcgactcttt acaggaaaat 840 gagttcagat tgtactacta taacaaattt aaagacgttg catcaacttt gaataaagcc 900 aaatctataa ttgggactac tgcgtccctg caatacatga aaaacgtgtt caaggagaag 960 taccttctgt cggaagatac atctggtaaa tttagcgttg acaagttaaa gttcgataaa 1020 ctgtacaaga tgttaacaga aatatatacc gaggataatt tcgtcaactt tttcaaagtc 1080 attaatcgga agacttactt aaacttcgat aaagccgtct tccgcattaa cattgtgcca 1140 gacgagaact atacaattaa ggatggtttc aatttgaaag gagctaactt aagcactaac 1200 tttaacggcc agaacacaga gatcaattca cgtaacttca caagacttaa gaattttaca 1260 gggctgttcg agttttataa gctgctttgc gtaagaggaa tcatcccctt tggattattt 1320 gacattatta agaagatcgc cgaaagtttt cgtgggtccg ttgttacgac aaacatttat 1380 ttaaattcga ca 1392 <210> 130 <211> 1383 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-BoNTs LC-His10-A2/DNA sequence 4 <400> 130 atgccgtttg tgaacaaaca attcaactat aaagatccgg taaacggagt tgacatcgcg 60 tacatcaaaa taccgaatgc tggtcagatg cagccagtaa aagctttcaa aatccacaac 120 aaaatctggg tcatccctga aagagatacc tttacaaatc ccgaggaggg ggacctgaat 180 ccaccaccag aggcgaaaca ggtcccggtt tcatactatg acagtaccta cctttctact 240 gataacgaga aagataacta tttgaagggt gtaactaaat tatttgaaag aatttacagc 300 acggatttgg gtcggatgct gttaacatca attgtgcgcg gcataccatt ctggggagga 360 tcgactatag acaccgagtt aaaggtgatt gatactaact gtataaatgt aatacaaccc 420 gacggttcat acagaagtga agagttaaat cttgtcatca ttggtccttc agctgatatt 480 atacagtttg aatgcaaaag ttttggacac gacgtactta acttgacgcg taatggttac 540 ggcagtacac agtacatacg cttcagccct gacttcacgt tcggtttcga ggaaagtctt 600 gaagttgata ctaaccccct gcttggcgca ggcaaattcg ccactgatcc cgctgtcaca 660 ctggctcacg aactgataca tgcagagcat agactttacg gcatcgcaat taacccaaac 720 cgggtcttta aggtaaatac gaatgcttac tacgaaatgt ctggcctgga agtatcattc 780 gaggagttga gaaccttcgg cggccacgac gctaaattta tcgacagtct tcaggaaaac 840 gagttccgtc tgtactacta caataaattt aaggatgtgg cttctacttt gaataaggcc 900 aaaagtatca tcgggactac tgcatccttg caatatatga agaacgtatt caaggaaaaa 960 tacttgttat cagaggacac gagtggcaag ttctctgttg acaagttaaa attcgataaa 1020 ttatacaaga tgttgactga gatatacaca gaggataact ttgtcaattt ttttaaggtt 1080 ataaatcgga agacctattt gaacttcgat aaggctgtct ttcgtattaa tatagtgccc 1140 gatgaaaact ataccatcaa agacggtttc aatctgaaag gtgccaattt gtcgaccaac 1200 ttcaacgggc aaaacacgga gatcaatagt agaaatttca cgcgtcttaa aaacttcact 1260 ggtctgttcg aattttataa gcttttgtgc gttcgcggaa taattccttt tcatcatcat 1320 catcaccatc atcaccacca tcgtgggtcc gtagtgacca ctaacatcta cttaaattct 1380 acc 1383 <210> 131 <211> 1422 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-BoNTs LC-His10-A2/DNA sequence 5 <400> 131 atggggttat tcgacataat caaaaagata gctgaatcct ttccgttcgt aaacaagcag 60 ttcaattata aagatccagt gaacggggtg gacattgcct acattaagat acccaatgcg 120 ggccagatgc aacccgtaaa agcgttcaaa atacacaaca aaatatgggt gattcctgag 180 cgcgacacat ttacgaaccc ggaagaaggt gatttgaatc cgccacctga agcgaaacag 240 gtccccgtga gttattacga tagcacgtac ttatcaactg ataatgagaa agataactat 300 ttgaagggcg tgacgaagtt attcgagcgc atatactcta ccgatcttgg tcggatgctg 360 ttaacgagca tcgtacgggg cattcccttc tggggtgggt ccacaatcga tacggagtta 420 aaagtcatcg acaccaactg tatcaacgtc atccagcctg acgggtcgta cagaagtgaa 480 gagttaaact tggttataat aggaccatcc gcagatatta tacaattcga gtgcaagtct 540 tttggtcatg atgttttgaa cctgacgcgc aacggatatg gctccacgca atacatccgc 600 ttctccccag attttacatt tggattcgaa gaatcattag aagtagacac aaacccactg 660 ctgggggctg gcaaattcgc aactgacccc gcagttacct tagcgcacga actgatacac 720 gcggaacatc ggctgtatgg catagcgatt aaccccaacc gtgttttcaa ggtaaacacg 780 aatgcgtact acgagatgtc tggtttggaa gtttcttttg aggagctgcg cacctttggg 840 ggtcatgatg cgaaatttat tgacagcttg caagagaatg agttcagatt atattactac 900 aataaattta aggatgtggc gagcactctg aataaggcaa aatccattat cggtactact 960 gcatctttgc agtatatgaa aaatgtgttc aaggagaagt acttgttgtc tgaggatacg 1020 agtggtaaat tctcagttga caaattgaaa tttgataagc tgtataagat gttgacagag 1080 atttatacgg aggataactt tgtgaatttc ttcaaagtca taaaccgtaa aacctacctg 1140 aattttgata aggcggtgtt tcggattaat atagttccgg acgagaatta tacaatcaag 1200 gacggcttca atttaaaagg agctaattta tccaccaatt ttaacggcca aaacacagaa 1260 atcaactctc gtaatttcac gcgtttaaag aacttcactg ggctgttcga attttataaa 1320 ttgttatgtg tccgcggcat aataccgttt catcatcacc atcatcatca tcaccatcac 1380 cgtggatcag tggtcaccac taatatttac ctgaacagta cg 1422 <210> 132 <211> 1422 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/DNA sequence 6 <400> 132 atgggtcttt tcgacataat aaagaaaatc gccgaaagct ttcatcacca tcaccatcat 60 catcatcacc atccgttcgt aaataagcag tttaattaca aagatcccgt gaacggcgtg 120 gacatcgcct atataaaaat cccgaatgcc ggccagatgc agcccgttaa ggccttcaag 180 atacacaata aaatctgggt cataccggag cgggatacat tcacaaaccc ggaagaaggt 240 gaccttaacc ctcccccgga ggcaaaacaa gttcccgttt catattatga ctctacgtac 300 ctttccaccg acaacgaaaa ggacaattat ctgaaaggcg tgacaaaact tttcgaacgc 360 atttattcta cagatcttgg tagaatgctg ttgacttcta ttgtcagagg aattcctttc 420 tggggcggga gcactataga cactgagtta aaggtaattg acacgaattg cataaatgtt 480 atacaaccgg acggcagtta ccgttcggag gagcttaatt tagttataat cggtccatca 540 gctgacataa ttcaatttga gtgcaaatca tttggacacg atgttttaaa tttgactcgt 600 aatggatacg gctccacgca atacatccgt ttcagtcctg attttacatt cggttttgaa 660 gagtctcttg aggtggacac gaaccctttg ttaggcgccg gcaaatttgc tactgatcca 720 gccgtcacat tagctcacga acttattcac gccgaacacc gtctgtacgg tattgcaata 780 aatccaaacc gggtctttaa agtaaacacg aacgcctact acgaaatgtc gggtcttgag 840 gtctcattcg aggaattaag aacctttgga ggtcacgatg cgaagttcat tgactccttg 900 caagagaacg aatttagact ttactattac aacaagttca aggatgtcgc gtcgacctta 960 aataaggcca aatccataat tgggacgaca gcaagccttc aatacatgaa aaacgtcttc 1020 aaggagaagt atcttttaag cgaggacacc agcggtaagt ttagtgtcga taagttaaaa 1080 ttcgacaaat tgtacaagat gcttactgag atttacacag aagacaattt tgttaatttt 1140 ttcaaagtta taaatcgtaa aacctatctg aatttcgata aagcggtttt ccgtatcaac 1200 attgtgccgg acgagaatta caccataaaa gacgggttca atttaaaggg cgcaaacctt 1260 tctaccaatt ttaacggaca aaatacggaa atcaactcac gcaattttac gcgcctgaaa 1320 aattttacag gcctttttga attttataag ttgctttgtg tacggggtat tatacccttt 1380 cggggatcag tggtgactac taatatatac ttgaactcga ct 1422 <210> 133 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> pET28b/Forward primer <400> 133 caccaccacc accaccactg agatc 25 <210> 134 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> pET28b/Backward primer <400> 134 ctcgctaccg cctccaccac tg 22 <210> 135 <211> 48 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC/Forward primer <400> 135 gtggtggtgg tggtggaagg gaatgatgcc acggacacaa agtaactt 48 <210> 136 <211> 53 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC/Backward primer <400> 136 ggaggcggta gcgagatgcc cttcgtcaac aaacagttca attataagga tcc 53 <210> 137 <211> 76 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-A2/Forward primer <400> 137 tgtgctgttc aaatagatgt ttgtggtcac aacgctcccg cgagagggtc tgtggtcacc 60 acaaatatat acctga 76 <210> 138 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-A2/Backward primer <400> 138 caccaccacc accaccactg agatc 25 <210> 139 <211> 72 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-Aurein 1.2-A2/Forward primer <400> 139 aaatgattct gcgatcttta ttatgatgtc aaacagtcca gagggtctgt ggtcaccaca 60 aatatatacc tg 72 <210> 140 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-Aurein 1.2-A2/Backward primer <400> 140 cgcgggagcg ttgtgaccac aaacat 26 <210> 141 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-His10-A2/ Forward primer <400> 141 gtgatggtga tggtgatggt gatggtggtg agagggtctg tggtcaccac aaatatatac 60 ctg 63 <210> 142 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-His10-A2/Backward primer <400> 142 cgcgggagcg ttgtgaccac aaacat 26 <210> 143 <211> 58 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-BoNTs LC-His10-A2/Forward primer <400> 143 aaatgattct gcgatcttta ttatgatgtc aaacagctcg ctaccgcctc caccactg 58 <210> 144 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-BoNTs LC-His10-A2/Backward primer <400> 144 atgcccttcg tcaacaaaca gttcaattat aaggatcc 38 <210> 145 <211> 52 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/Backward primer <400> 145 gtgatggtga tggtgatggt gatggtggtg ctcgctaccg cctccaccac tg 52 <210> 146 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/Backward primer <400> 146 atgcccttcg tcaacaaaca gttcaattat aaggatcc 38 <210> 147 <211> 61 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/Forward primer <400> 147 aaatgattct gcgatcttta ttatgatgtc aaacagtccc tcgctaccgc ctccaccact 60 g 61 <210> 148 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/Backward primer <400> 148 caccaccatc accatcacca tcaccatc 28 <210> 149 <211> 427 <212> PRT <213> Artificial Sequence <220> <223> RBD <400> 149 Met Lys Asn Ile Ile Asn Thr Ser Ile Leu Asn Leu Arg Tyr Glu Ser 1 5 10 15 Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile Gly 20 25 30 Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu Phe 35 40 45 Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile Val 50 55 60 Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg Ile 65 70 75 80 Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile Ile 85 90 95 Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr Gly 100 105 110 Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg Val 115 120 125 Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn Arg 130 135 140 Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys Ile 145 150 155 160 Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu Gly 165 170 175 Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys Arg 180 185 190 Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp Lys 195 200 205 Glu Leu Asn Glu Lys Glu Ile Lys Asp Leu Tyr Asp Asn Gln Ser Asn 210 215 220 Ser Gly Ile Leu Lys Asp Phe Trp Gly Asp Tyr Leu Gln Tyr Asp Lys 225 230 235 240 Pro Tyr Tyr Met Leu Asn Leu Tyr Asp Pro Asn Lys Tyr Ile Asp Val 245 250 255 Asn Asn Val Gly Ile Arg Gly Tyr Met Tyr Leu Lys Gly Pro Arg Gly 260 265 270 Asn Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser Leu Tyr Met Gly 275 280 285 Thr Lys Phe Ile Ile Lys Lys Tyr Ala Ser Gly Asn Lys Asp Asn Ile 290 295 300 Val Arg Asn Asn Asp Arg Val Tyr Ile Asn Val Val Val Lys Asn Lys 305 310 315 320 Glu Tyr Arg Leu Ala Thr Asn Ala Ser Gln Ala Gly Val Glu Lys Ile 325 330 335 Leu Ser Ala Leu Glu Ile Pro Asp Val Gly Asn Leu Ser Gln Val Val 340 345 350 Val Met Lys Ser Lys Asn Asp Gln Gly Ile Thr Asn Lys Cys Lys Met 355 360 365 Asn Leu Gln Asp Asn Asn Gly Asn Asp Ile Gly Phe Ile Gly Phe His 370 375 380 Gln Phe Asn Asn Ile Ala Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg 385 390 395 400 Gln Ile Glu Arg Ser Ser Arg Thr Leu Gly Cys Ser Trp Glu Phe Ile 405 410 415 Pro Val Asp Asp Gly Trp Gly Glu Arg Pro Leu 420 425 <210> 150 <211> 427 <212> PRT <213> Artificial Sequence <220> <223> RBD' <400> 150 Met Lys Asn Ile Ile Asn Thr Ser Ile Leu Asn Leu Arg Tyr Glu Ser 1 5 10 15 Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile Gly 20 25 30 Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu Phe 35 40 45 Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile Val 50 55 60 Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg Ile 65 70 75 80 Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile Ile 85 90 95 Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr Gly 100 105 110 Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg Val 115 120 125 Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn Arg 130 135 140 Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys Ile 145 150 155 160 Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu Gly 165 170 175 Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys Arg 180 185 190 Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp Lys 195 200 205 Glu Leu Asn Glu Lys Glu Ile Lys Asp Leu Tyr Asp Asn Gln Ser Asn 210 215 220 Ser Gly Ile Leu Lys Asp Phe Trp Gly Asp Tyr Leu Gln Tyr Asp Lys 225 230 235 240 Pro Tyr Tyr Met Leu Asn Leu Tyr Asp Pro Asn Lys Tyr Val Asp Val 245 250 255 Asn Asn Val Gly Ile Arg Gly Tyr Met Tyr Leu Lys Gly Pro Arg Gly 260 265 270 Ser Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser Leu Tyr Arg Gly 275 280 285 Thr Lys Phe Ile Ile Lys Lys Tyr Ala Ser Gly Asn Lys Asp Asn Ile 290 295 300 Val Arg Asn Asn Asp Arg Val Tyr Ile Asn Val Val Val Lys Asn Lys 305 310 315 320 Glu Tyr Arg Leu Ala Thr Asn Ala Ser Gln Ala Gly Val Glu Lys Ile 325 330 335 Leu Ser Ala Leu Glu Ile Pro Asp Val Gly Asn Leu Ser Gln Val Val 340 345 350 Val Met Lys Ser Lys Asn Asp Gln Gly Ile Thr Asn Lys Cys Lys Met 355 360 365 Asn Leu Gln Asp Asn Asn Gly Asn Asp Ile Gly Phe Ile Gly Phe His 370 375 380 Gln Phe Asn Asn Ile Ala Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg 385 390 395 400 Gln Ile Glu Arg Ser Ser Arg Thr Leu Gly Cys Ser Trp Glu Phe Ile 405 410 415 Pro Val Asp Asp Gly Trp Gly Glu Arg Pro Leu 420 425 <210> 151 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> A2 <400> 151 Arg Gly Asn Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser 1 5 10 <210> 152 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> A2' <400> 152 Arg Gly Ser Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser 1 5 10 <210> 153 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> SV2C <400> 153 Asp Val Ile Lys Pro Leu Gln Ser Asp Glu Tyr Ala Leu Leu Thr Arg 1 5 10 15 Asn Val Glu Arg Asp Lys Tyr Ala Asn Phe Thr Ile Asn Phe Thr Met 20 25 30 Glu Asn Gln Ile His Thr Gly Met Glu Tyr Asp Asn Gly Arg Phe Ile 35 40 45 Gly Val Lys Phe Lys Ser Val Thr Phe Lys Asp Ser Val Phe Lys Ser 50 55 60 Cys Thr Phe Glu Asp Val Thr Ser Val Asn Thr Tyr Phe Lys Asn Cys 65 70 75 80 Thr Phe Ile Asp Thr Val Phe Asp Asn Thr Asp Phe Glu Pro Tyr Lys 85 90 95 Phe Ile Asp Ser Glu Phe Lys Asn Cys Ser Phe Phe His Asn Lys Thr 100 105 110 Gly Cys Gln Ile Thr Phe Asp 115 <210> 154 <211> 1281 <212> DNA <213> Artificial Sequence <220> <223> RBD <400> 154 atgaaaaaca ttattaatac ttccatactt aacctgcgct acgagtctaa ccatttgatt 60 gacttatcgc ggtacgcttc taaaataaac atcgggagca aggtaaactt tgatccgata 120 gataaaaacc agattcaatt gttcaactta gagtctagta aaatagaggt tattttaaaa 180 aatgccattg tttataactc gatgtacgaa aatttctcga cctcgttttg gatacggata 240 ccgaagtact ttaactctat ttccttgaat aatgagtaca ccatcataaa ttgcatggaa 300 aacaattccg gttggaaagt gtcattaaac tatggcgaga taatttggac tttgcaggac 360 actcaggaga taaagcagcg tgtagtcttt aagtattccc agatgataaa catctcggac 420 tacataaatc gttggatttt tgtaaccata acaaataatc gccttaacaa tagcaaaatt 480 tatatcaacg gccggttgat cgatcaaaaa ccaatttcca acttggggaa catacacgct 540 tcgaataaca tcatgtttaa gcttgacggt tgtagagata cgcatcgcta catttggata 600 aagtatttta acctgtttga caaggagttg aatgagaaag aaattaagga cttatatgat 660 aatcaatcaa acagtgggat attgaaagat ttctggggcg actatcttca gtacgataag 720 ccctattata tgttgaactt atacgaccca aacaaataca ttgacgtaaa caatgtgggt 780 atcagaggat atatgtactt aaagggccca agagggaatg taatgacgac gaacatatac 840 cttaacagca gtctttacat gggcacgaaa ttcataataa aaaaatatgc gtccgggaat 900 aaggacaata ttgtaagaaa taacgaccgt gtatatatca atgtcgtggt caaaaacaaa 960 gagtaccggc tggctaccaa tgcttcccag gcaggtgtcg agaaaatact ttcggcgctt 1020 gaaattccag acgtgggcaa tctgtcacaa gtggtcgtta tgaaaagtaa aaatgatcaa 1080 gggatcacca ataagtgcaa gatgaatttg caagacaata atggtaacga tataggattt 1140 atagggtttc accagttcaa taacatcgct aagctggtcg ctagtaactg gtataataga 1200 caaattgaac ggtccagtcg cacgcttggc tgttcttggg agttcattcc ggttgatgat 1260 ggatggggcg aacgcccact g 1281 <210> 155 <211> 1281 <212> DNA <213> Artificial Sequence <220> <223> RBD' <400> 155 atgaaaaaca ttattaatac ttccatactt aacctgcgct acgagtctaa ccatttgatt 60 gacttatcgc ggtacgcttc taaaataaac atcgggagca aggtaaactt tgatccgata 120 gataaaaacc agattcaatt gttcaactta gagtctagta aaatagaggt tattttaaaa 180 aatgccattg tttataactc gatgtacgaa aatttctcga cctcgttttg gatacggata 240 ccgaagtact ttaactctat ttccttgaat aatgagtaca ccatcataaa ttgcatggaa 300 aacaattccg gttggaaagt gtcattaaac tatggcgaga taatttggac tttgcaggac 360 actcaggaga taaagcagcg tgtagtcttt aagtattccc agatgataaa catctcggac 420 tacataaatc gttggatttt tgtaaccata acaaataatc gccttaacaa tagcaaaatt 480 tatatcaacg gccggttgat cgatcaaaaa ccaatttcca acttggggaa catacacgct 540 tcgaataaca tcatgtttaa gcttgacggt tgtagagata cgcatcgcta catttggata 600 aagtatttta acctgtttga caaggagttg aatgagaaag aaattaagga cttatatgat 660 aatcaatcaa acagtgggat attgaaagat ttctggggcg actatcttca gtacgataag 720 ccctattata tgttgaactt atacgaccca aacaaatacg tggacgtaaa caatgtgggt 780 atcagaggat atatgtactt aaagggccca agagggtctg taatgacgac gaacatatac 840 cttaacagca gtctttaccg tggcacgaaa ttcataataa aaaaatatgc gtccgggaat 900 aaggacaata ttgtaagaaa taacgaccgt gtatatatca atgtcgtggt caaaaacaaa 960 gagtaccggc tggctaccaa tgcttcccag gcaggtgtcg agaaaatact ttcggcgctt 1020 gaaattccag acgtgggcaa tctgtcacaa gtggtcgtta tgaaaagtaa aaatgatcaa 1080 gggatcacca ataagtgcaa gatgaatttg caagacaata atggtaacga tataggattt 1140 atagggtttc accagttcaa taacatcgct aagctggtcg ctagtaactg gtataataga 1200 caaattgaac ggtccagtcg cacgcttggc tgttcttggg agttcattcc ggttgatgat 1260 ggatggggcg aacgcccact g 1281 <210> 156 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> A2 <400> 156 agagggaatg taatgacgac gaacatatac cttaacagca gt 42 <210> 157 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> A2' <400> 157 gggtctgtaa tgacgacgaa catatacctt aacagcagtc tt 42 <210> 158 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> SV2C <400> 158 gacgtgataa agccccttca gtcagatgaa tatgcattgc tgactcgcaa cgtggaacgt 60 gacaagtatg ccaattttac cataaatttc acaatggaaa accagatcca tacgggcatg 120 gagtacgaca acggtcggtt tatcggggta aaattcaagt cagtaacctt taaggattcg 180 gtgttcaaaa gttgcacatt tgaggacgtt acctccgtaa acacctactt caaaaattgc 240 acttttattg atacagtatt cgacaacacc gatttcgagc catataaatt catcgacagt 300 gagttcaaga actgcagttt ctttcataat aagaccggct gccagattac tttcgat 357 <210> 159 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> pET28b-EGFP/Forward primer <400> 159 gctaccgcct ccaccgtggt g 21 <210> 160 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> pET28b-EGFP/Backward primer <400> 160 tgagatccgg ctgctaacaa agcccgaaa 29 <210> 161 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> RBD insert/Forward primer <400> 161 agcagccgga tctcacagtg ggcgttcgcc ccatccat 38 <210> 162 <211> 62 <212> DNA <213> Artificial Sequence <220> <223> RBD insert/Backward primer <400> 162 ggtggaggcg gtagcaaaaa cattattaat acttccatac ttaacctgcg ctacgagtct 60 aa 62 <210> 163 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM1/Forward primer <400> 163 tatgaatttc gtgccacggt aaagactgct gtt 33 <210> 164 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM1/Backward primer <400> 164 aacagcagtc tttaccgtgg cacgaaattc ata 33 <210> 165 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM2/Forward primer <400> 165 ttcgtcgtca ttacagaccc tcttgggccc tt 32 <210> 166 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM2/Backward primer <400> 166 aagggcccaa gagggtctgt aatgacgacg aa 32 <210> 167 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM3/Forward primer <400> 167 cacattgttt acgtccacgt atttgtttgg gtc 33 <210> 168 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM3/Backward primer <400> 168 gacccaaaca aaacgtggac gtaaacaatg tg 32 <210> 169 <211> 86 <212> DNA <213> Artificial Sequence <220> <223> EGFP-A2/Forward primer <400> 169 ggtggaggcg gtagcagagg gaatgtaatg acgacgaaca tataccttaa cagcagttga 60 gatccggctg ctaacaaagc ccgaaa 86 <210> 170 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> EGFP-A2/Backward primer <400> 170 gtggtggtgg tggtggtgct cg 22 <210> 171 <211> 75 <212> DNA <213> Artificial Sequence <220> <223> EGFP-Aurein1.2-A2/Forward primer <400> 171 gctaccgcct ccaccggcct gtttgacatc atcaaaaaaa tcgccgaaag ctttgctacc 60 gcctccaccg tggtg 75 <210> 172 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> EGFP-Aurein1.2-A2/Backward primer <400> 172 agagggaatg taatgacgac gaacatatac cttaacag 38 <210> 173 <211> 75 <212> DNA <213> Artificial Sequence <220> <223> EGFP-Aurein1.2-RBD/Forward primer <400> 173 gctaccgcct ccaccggcct gtttgacatc atcaaaaaaa tcgccgaaag ctttgctacc 60 gcctccaccg tggtg 75 <210> 174 <211> 44 <212> DNA <213> Artificial Sequence <220> <223> EGFP-Aurein1.2-RBD/Backward primer <400> 174 gactcgtagc gcaggttaag tatggaagta ttaataatgt tttt 44 <110> Research and Business Foundation SungKyunKwan University <120> A composition comprising neuro-targeted proteins <130> MP19-178 <150> KR 10-2018-0101615 <151> 2018-08-28 <160> 174 <170> KoPatentIn 3.0 <210> 1 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 <400> 1 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Lys Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Lys Gly Tyr Asn Lys 435 440 445 <210> 2 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype <400> 2 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Val Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Lys Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Lys Gly Tyr Asn Lys 435 440 445 <210> 3 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 3 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 4 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype <400> 4 Met Gln Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Val Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Lys Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Lys Gly Tyr Asn Lys 435 440 445 <210> 5 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A5 subtype <400> 5 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Glu Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Glu His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Glu Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 6 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 6 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Ile Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 7 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A5 subtype <400> 7 Met Leu Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Glu Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Glu His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Glu Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 8 <211> 444 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A3 subtype <400> 8 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Arg Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Glu Gly Val Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Ile Lys Leu Phe Asp 85 90 95 Arg Ile Tyr Ser Thr Gly Leu Gly Arg Met Leu Leu Ser Phe Ile Val 100 105 110 Lys Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Glu Pro Gly Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Thr Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Phe Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Thr Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Ala His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Leu Lys Val Lys Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly Asn Asp Thr Asn 260 265 270 Phe Ile Asp Ser Leu Trp Gln Lys Lys Phe Ser Arg Asp Ala Tyr Asp 275 280 285 Asn Leu Gln Asn Ile Ala Arg Ile Leu Asn Glu Ala Lys Thr Ile Val 290 295 300 Gly Thr Thr Thr Pro Leu Gln Tyr Met Lys Asn Ile Phe Ile Arg Lys 305 310 315 320 Tyr Phe Leu Ser Glu Asp Ala Ser Gly Lys Ile Ser Val Asn Lys Pro 325 330 335 Ala Phe Lys Glu Phe Tyr Arg Val Leu Thr Arg Gly Phe Thr Glu Leu 340 345 350 Glu Phe Val Asn Pro Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Asn Glu Gly Phe Asn Leu Glu Gly Ala Asn Ser Asn Gly Gln 385 390 395 400 Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu Lys Asn Phe Thr 405 410 415 Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg Gly Ile Ile Pro 420 425 430 Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 <210> 9 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 9 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Leu Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 10 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 10 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Ile Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 11 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A <400> 11 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Ile Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Asn Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Ile Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Glu Val Ala Ser Ile Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Arg Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Met Asn Ile Val Pro Glu Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 12 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 12 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Thr Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Lys Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asn Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 13 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A4 subtype <400> 13 Met Pro Leu Val Asn Gln Gln Ile Asn Tyr Tyr Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Lys Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Val Trp Val Ile Pro Glu Arg 35 40 45 Asp Ile Phe Thr Asn Pro Glu Glu Val Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Ile Ser Tyr Tyr Asp Ser Ala Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Ile Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Ile Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Gly Lys Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Ile Ile Gln Leu Asp Asp Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Ala Ile Ile Gly Pro Ser Ala Asn Ile 145 150 155 160 Ile Glu Ser Gln Cys Ser Ser Phe Arg Asp Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Val Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Gln Asp Pro Ala Val Ala Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Thr Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ala Gly Leu 245 250 255 Glu Val Ser Leu Glu Glu Leu Ile Thr Phe Gly Gly Asn Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Lys Lys Glu Phe Ser Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Ala Thr Gly Lys Phe Leu Val Asp Arg Leu 325 330 335 Lys Phe Asp Glu Leu Tyr Lys Leu Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Asp Val Asn Tyr 370 375 380 Thr Ile His Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Ile Glu Ile Asn Asn Lys Asn Phe Asp Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 14 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype <400> 14 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Glu Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 15 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype <400> 15 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Val 20 25 30 Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg Asp 35 40 45 Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu Ala 50 55 60 Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr Asp 65 70 75 80 Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu Arg 85 90 95 Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val Arg 100 105 110 Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys Val 115 120 125 Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr Arg 130 135 140 Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile Ile 145 150 155 160 Gln Phe Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn Leu Thr Arg 165 170 175 Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe Thr 180 185 190 Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu Gly 195 200 205 Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu Leu 210 215 220 Ile His Ala Gly His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn Arg 225 230 235 240 Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu Glu 245 250 255 Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys Phe 260 265 270 Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn Lys 275 280 285 Phe Lys Asp Ile Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Val Gly 290 295 300 Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys Tyr 305 310 315 320 Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu Lys 325 330 335 Phe Asp Lys Leu Tyr Lys Met Lys Thr Glu Ile Tyr Thr Glu Asp Asp 340 345 350 Asn Phe Val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Lys Ile Asn Ile Val Pro Lys Val Asn Tyr 370 375 380 Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Thr Ser Lys Thr Lys Ser Leu Asp Glu Gly Tyr Asn Lys 435 440 445 <210> 16 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 16 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asp Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Phe Gln Thr Leu Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Thr Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asn Lys Leu Tyr Lys Ser Leu 340 345 350 Met Leu Gly Phe Thr Glu Ile Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Gly Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 17 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B-B1 subtype <400> 17 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 18 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 18 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asp Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Gln Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Thr Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asn Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Ile Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Gly Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 19 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 19 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Met Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asn Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 20 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 20 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asp Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Gln Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Thr Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asn Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Gly Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 21 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 21 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Arg 435 440 <210> 22 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 22 Met Ser Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asp Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asn Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 23 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 23 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asp Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 24 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B-B8 subtype <400> 24 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Gly Glu Glu Arg Lys Glu Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asp Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Gly Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Arg 435 440 <210> 25 <211> 439 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B-B1 subtype <400> 25 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Ile Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Met Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Leu Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Trp Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Ala Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Ile Val Asn Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Phe Val Glu Asp Ser Glu Gly Lys Tyr 325 330 335 Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu Met Phe 340 345 350 Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys Thr Arg 355 360 365 Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys Asn Leu 370 375 380 Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile Ser Asp 385 390 395 400 Lys Asn Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile Asn Lys 405 410 415 Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Ile Tyr Lys Ile 420 425 430 Gln Met Cys Lys Ser Val Lys 435 <210> 26 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B-B8 subtype <400> 26 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Gly Glu Glu Arg Lys Glu Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Gly Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asp Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asn Met Gly Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Arg 435 440 <210> 27 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 27 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Trp Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Ala Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Ile Val Asn Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Ser Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Asn Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 28 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/B <400> 28 Met Pro Val Thr Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn 1 5 10 15 Asn Asn Ile Ile Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg 20 25 30 Tyr Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp Ile Ile Pro Glu 35 40 45 Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 55 60 Ile Phe Asn Arg Asp Val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn 65 70 75 80 Thr Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met Ile Lys Leu Phe 85 90 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile 100 105 110 Ile Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu 115 120 125 Phe Asn Thr Asn Ile Ala Ser Val Thr Val Asn Lys Leu Ile Ser Asn 130 135 140 Pro Gly Glu Val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu Ile Ile 145 150 155 160 Phe Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly 165 170 175 Ile Gln Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gln 180 185 190 Met Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu 195 200 205 Asn Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro 210 215 220 Ala Leu Ile Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr 225 230 235 240 Gly Ile Lys Val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe 245 250 255 Phe Met Gln Ser Thr Asp Ala Ile Gln Ala Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Gln Asp Pro Ser Ile Ile Thr Pro Ser Thr Asp Lys Ser Ile 275 280 285 Tyr Asp Lys Val Leu Gln Asn Phe Arg Gly Ile Val Asp Arg Leu Asn 290 295 300 Lys Val Leu Val Cys Ile Ser Asp Pro Ser Ile Asn Ile Asn Ile Tyr 305 310 315 320 Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly 325 330 335 Lys Tyr Ser Ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu 340 345 350 Met Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys 355 360 365 Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Val Lys Ile Lys 370 375 380 Asn Leu Leu Asp Asn Glu Ile Tyr Thr Ile Glu Glu Gly Phe Asn Ile 385 390 395 400 Ser Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile 405 410 415 Asn Lys Gln Ala Tyr Glu Glu Ile Ser Lys Glu His Leu Ala Val Tyr 420 425 430 Lys Ile Gln Met Cys Lys Ser Val Lys 435 440 <210> 29 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/C1 <400> 29 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Thr Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asn Ser Asn Pro Asn Leu Asn Lys Pro Arg Val 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Asp Lys Asp Pro Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ser Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Thr Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Thr Val Asn Arg Asn Lys Phe Val Glu Leu Tyr Asn 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 30 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/C1 <400> 30 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Ile Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asp Ser Asn Pro Asn Leu Lys Lys Pro Arg Val 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Glu Lys Asp Thr Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ala Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Arg Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Glu 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Ala Val Asp Arg Asn Lys Phe Ala Glu Leu Tyr Lys 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 31 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/C1 <400> 31 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Ile Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asp Ser Asn Pro Asn Leu Asn Lys Pro Arg Val 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Glu Lys Asp Thr Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ala Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asn 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Thr Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Arg Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Lys Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Gly Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Ala Val Asp Arg Asn Lys Phe Ala Glu Leu Tyr Lys 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 32 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/C1 <400> 32 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Ile Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asp Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Xaa 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Glu Lys Asp Thr Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ala Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asn 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Arg Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Lys Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Ala Val Asp Arg Asn Lys Phe Ala Glu Leu Tyr Lys 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Ala Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 33 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/CD <400> 33 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Ile Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asp Ser Asn Pro Asn Leu Asn Lys Pro Arg Val 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Glu Lys Asp Thr Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ala Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asn 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Arg Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Ala Val Asp Arg Asn Lys Phe Ala Glu Leu Tyr Lys 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 34 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> BoNT/CD <400> 34 Met Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro Val Asp Asn 1 5 10 15 Lys Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu 20 25 30 Pro Glu Lys Ala Phe Arg Ile Ile Gly Asn Ile Trp Val Ile Pro Asp 35 40 45 Arg Phe Ser Arg Asp Ser Asn Pro Asn Leu Asn Lys Pro Arg Val 50 55 60 Thr Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp 65 70 75 80 Ser Glu Lys Asp Thr Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ala Thr 100 105 110 Asp Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp 115 120 125 Phe Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn 130 135 140 Asn Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly 145 150 155 160 Pro Arg Glu Asn Ile Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr 165 170 175 Asn Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile 180 185 190 Ser Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asn 195 200 205 Val Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile 210 215 220 Leu Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly 225 230 235 240 Ile Ala Ile Pro Asn Asp Gln Arg Ile Ser Ser Val Thr Ser Asn Ile 245 250 255 Phe Tyr Ser Gln Tyr Lys Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala 260 265 270 Phe Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr 275 280 285 Phe Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu 290 295 300 Asn Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly 305 310 315 320 Glu Tyr Lys Gln Lys Leu Ile Arg Lys Tyr Arg Phe Val Val Glu Ser 325 330 335 Ser Gly Glu Val Ala Val Asp Arg Asn Lys Phe Ala Glu Leu Tyr Lys 340 345 350 Glu Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn 355 360 365 Val Gln Asn Arg Lys Ile Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr 370 375 380 Ala Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn 385 390 395 400 Ile Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser 405 410 415 Arg Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu 420 425 430 Phe Thr Lys Phe Cys His Lys Ala Ile Asp Gly Arg Ser Leu Tyr Asn 435 440 445 Lys <210> 35 <211> 442 <212> PRT <213> Artificial Sequence <220> <223> BoNT/D <400> 35 Met Thr Trp Pro Val Lys Asp Phe Asn Tyr Ser Asp Pro Val Asn Asp 1 5 10 15 Asn Asp Ile Leu Tyr Leu Arg Ile Pro Gln Asn Lys Leu Ile Thr Thr 20 25 30 Pro Val Lys Ala Phe Met Ile Thr Gln Asn Ile Trp Val Ile Pro Glu 35 40 45 Arg Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro 50 55 60 Thr Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp 65 70 75 80 Glu Gln Lys Asp Thr Phe Leu Lys Gly Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Glu Arg Asp Ile Gly Lys Lys Leu Ile Asn Tyr Leu Val Val 100 105 110 Gly Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp 115 120 125 Phe Thr Arg His Thr Thr Asn Ile Ala Val Glu Lys Phe Glu Asn Gly 130 135 140 Ser Trp Lys Val Thr Asn Ile Ile Thr Pro Ser Val Leu Ile Phe Gly 145 150 155 160 Pro Leu Pro Asn Ile Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly 165 170 175 Gln Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu 180 185 190 Lys Val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr Ser Asn 195 200 205 Gln Ser Ser Ala Val Leu Gly Lys Ser Ile Phe Cys Met Asp Pro Val 210 215 220 Ile Ala Leu Met His Glu Leu Thr His Ser Leu His Gln Leu Tyr Gly 225 230 235 240 Ile Asn Ile Pro Ser Asp Lys Arg Ile Arg Pro Gln Val Ser Glu Gly 245 250 255 Phe Phe Ser Gln Asp Gly Pro Asn Val Gln Phe Glu Glu Leu Tyr Thr 260 265 270 Phe Gly Gly Leu Asp Val Glu Ile Ile Pro Gln Ile Glu Arg Ser Gln 275 280 285 Leu Arg Glu Lys Ala Leu Gly His Tyr Lys Asp Ile Ala Lys Arg Leu 290 295 300 Asn Asn Ile Asn Lys Thr Ile Pro Ser Ser Trp Ile Ser Asn Ile Asp 305 310 315 320 Lys Tyr Lys Lys Ile Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn 325 330 335 Thr Gly Asn Phe Val Val Asn Ile Asp Lys Phe Asn Ser Leu Tyr Ser 340 345 350 Asp Leu Thr Asn Val Met Ser Glu Val Val Tyr Ser Ser Gln Tyr Asn 355 360 365 Val Lys Asn Arg Thr His Tyr Phe Ser Arg His Tyr Leu Pro Val Phe 370 375 380 Ala Asn Ile Leu Asp Asp Asn Ile Tyr Thr Ile Arg Asp Gly Phe Asn 385 390 395 400 Leu Thr Asn Lys Gly Phe Asn Ile Glu Asn Ser Gly Gln Asn Ile Glu 405 410 415 Arg Asn Pro Ala Leu Gln Lys Leu Ser Ser Ser Glu Ser Val Val Asp Leu 420 425 430 Phe Thr Lys Val Cys Leu Arg Leu Thr Lys 435 440 <210> 36 <211> 442 <212> PRT <213> Artificial Sequence <220> <223> BoNT/D <400> 36 Met Thr Trp Pro Val Lys Asp Phe Asn Tyr Ser Asp Pro Val Asn Asp 1 5 10 15 Asn Asp Ile Leu Tyr Leu Arg Ile Pro Gln Asn Lys Leu Ile Thr Thr 20 25 30 Pro Val Lys Ala Phe Met Ile Thr Gln Asn Ile Trp Val Ile Pro Glu 35 40 45 Arg Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro 50 55 60 Thr Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp 65 70 75 80 Glu Gln Lys Asp Thr Phe Leu Lys Gly Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Glu Arg Asp Ile Gly Lys Lys Leu Ile Asn Tyr Leu Val Val 100 105 110 Gly Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp 115 120 125 Phe Thr Arg His Thr Thr Asn Ile Ala Val Glu Lys Phe Glu Asn Gly 130 135 140 Ser Trp Lys Val Thr Asn Ile Ile Thr Pro Ser Val Leu Ile Phe Gly 145 150 155 160 Pro Leu Pro Asn Ile Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly 165 170 175 Gln Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu 180 185 190 Lys Val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr Ser Asn 195 200 205 Gln Ser Ser Ala Val Leu Gly Lys Ser Ile Phe Cys Met Asp Pro Val 210 215 220 Ile Ala Leu Met His Glu Leu Thr His Ser Leu His Gln Leu Tyr Gly 225 230 235 240 Ile Asn Ile Pro Ser Asp Lys Arg Ile Arg Pro Gln Val Ser Glu Gly 245 250 255 Phe Phe Ser Gln Asp Gly Pro Asn Val Gln Phe Glu Glu Leu Tyr Thr 260 265 270 Phe Gly Gly Ser Asp Val Glu Ile Ile Pro Gln Ile Glu Arg Ser Gln 275 280 285 Leu Arg Glu Lys Ala Leu Asp His Tyr Lys Asp Ile Ala Lys Arg Leu 290 295 300 Asn Asn Ile Asn Lys Thr Ile Pro Ser Ser Trp Ser Ser Asn Ile Asp 305 310 315 320 Lys Tyr Lys Lys Ile Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn 325 330 335 Thr Gly Asn Phe Val Val Ser Ile Asp Lys Phe Asn Arg Leu Tyr Ser 340 345 350 Asp Leu Thr Asn Val Met Ser Glu Val Val Tyr Ser Ser Gln Tyr Asn 355 360 365 Val Lys Asn Arg Thr His Tyr Phe Ser Lys Arg Tyr Leu Pro Val Phe 370 375 380 Ala Asn Ile Leu Asp Asp Asn Ile Tyr Thr Ile Ile Asp Gly Phe Asn 385 390 395 400 Leu Thr Thr Lys Gly Phe Asn Ile Glu Asn Ser Gly Gln Asn Ile Glu 405 410 415 Arg Asn Pro Ala Leu Gln Lys Leu Ser Ser Ser Glu Ser Val Val Asp Leu 420 425 430 Phe Thr Lys Val Cys Leu Arg Leu Thr Lys 435 440 <210> 37 <211> 442 <212> PRT <213> Artificial Sequence <220> <223> BoNT/D <400> 37 Met Thr Trp Pro Val Lys Asp Phe Asn Tyr Ser Asp Pro Val Asn Asp 1 5 10 15 Asn Asp Ile Leu Tyr Leu Arg Ile Pro Gln Asn Lys Leu Ile Thr Thr 20 25 30 Pro Val Lys Ala Phe Met Ile Thr Gln Asn Ile Trp Val Ile Pro Glu 35 40 45 Arg Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro 50 55 60 Thr Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp 65 70 75 80 Glu Gln Lys Asp Thr Phe Leu Lys Gly Ile Ile Lys Leu Phe Lys Arg 85 90 95 Ile Asn Glu Arg Asp Ile Gly Lys Lys Leu Ile Asn Tyr Leu Val Val 100 105 110 Gly Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp 115 120 125 Phe Thr Arg His Thr Thr Asn Ile Ala Val Glu Lys Phe Glu Asn Gly 130 135 140 Ser Trp Lys Val Thr Asn Ile Ile Thr Pro Ser Val Leu Ile Phe Gly 145 150 155 160 Pro Leu Pro Asn Ile Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly 165 170 175 Gln Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu 180 185 190 Lys Val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr Ser Asn 195 200 205 Gln Ser Ser Ala Val Leu Gly Lys Ser Ile Phe Cys Met Asp Pro Val 210 215 220 Ile Ala Leu Met His Glu Leu Thr His Ser Leu His Gln Leu Tyr Gly 225 230 235 240 Ile Asn Ile Pro Ser Asp Lys Arg Ile Arg Pro Gln Val Ser Glu Gly 245 250 255 Phe Phe Ser Gln Asp Gly Pro Asn Val Gln Phe Glu Glu Leu Tyr Thr 260 265 270 Phe Gly Gly Ser Asp Val Glu Ile Ile Pro Gln Ile Glu Arg Leu Gln 275 280 285 Leu Arg Glu Lys Ala Leu Gly His Tyr Lys Asp Ile Ala Lys Arg Leu 290 295 300 Asn Asn Ile Asn Lys Thr Ile Pro Ser Ser Trp Ser Ser Asn Ile Asp 305 310 315 320 Lys Tyr Lys Lys Ile Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn 325 330 335 Thr Gly Asn Phe Val Val Asn Ile Asp Lys Phe Asn Ser Leu Tyr Ser 340 345 350 Asp Leu Thr Asn Val Met Ser Glu Val Val Tyr Ser Ser Gln Tyr Asn 355 360 365 Val Lys Asn Arg Thr His Tyr Phe Ser Lys His Tyr Leu Pro Val Phe 370 375 380 Ala Asn Ile Leu Asp Asp Asn Ile Tyr Thr Ile Ile Asn Gly Phe Asn 385 390 395 400 Leu Thr Thr Lys Gly Phe Asn Ile Glu Asn Ser Gly Gln Asn Ile Glu 405 410 415 Arg Asn Pro Ala Leu Gln Lys Leu Ser Ser Ser Glu Ser Val Val Asp Leu 420 425 430 Phe Thr Lys Val Cys Leu Arg Leu Thr Arg 435 440 <210> 38 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 38 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Asp Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Arg Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Cys Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Arg Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 39 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 39 Met Pro Thr Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asn Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Gln Glu Lys 65 70 75 80 Asp Lys Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Arg Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Gly Asp Phe Ile Ile Asn Asp 115 120 125 Ala Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Lys Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 40 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E3 subtype <400> 40 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln His Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Ile Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Thr Cys Ile Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Arg Lys Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Asn Asp Leu Asn Ile Ile Thr Val Ala Gln Tyr Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asn Asp Tyr Arg Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asp Asp Ile Leu Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Lys Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 41 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E1 subtype <400> 41 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Asp Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 42 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 42 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Lys 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Arg Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Asn Gln Ser Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Arg Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Tyr Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asn Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asn Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Thr Asn Leu Asn Pro Arg Ile Ile Thr Gln Leu Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Phe Ser Lys Gly Ile Thr 420 <210> 43 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 43 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ile Gln Asp Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 44 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 44 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln His Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Ile Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Thr Cys Ile Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Arg Lys Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Asn Asp Leu Asn Ile Ile Thr Val Ala Gln Tyr Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asn Asp Tyr Arg Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Lys Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 45 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E10 subtype <400> 45 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Lys 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asn Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Thr Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Lys Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Asn Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Gln 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asn Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asn Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Thr Asn Leu Asn Pro Arg Ile Ile Thr Gln Leu Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Phe Ser Lys Gly Ile Thr 420 <210> 46 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 46 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Asn 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asn Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Asn Asp Gln Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Val Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Arg Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Arg Asp Asp Asp Phe Ile Ile Asn Asp 115 120 125 Gly Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Thr Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Val Ser Leu Ile Asn Asn Tyr Ser Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Ile His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Asn Asn Leu Asn Ile Ile Thr Ser Ser Gln Leu Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asp Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Val Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asn Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Asn Val Asn Phe 370 375 380 Arg Gly Gln Asn Pro Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Asp Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 47 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 47 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Asp Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Gln Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 48 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 48 Met Pro Thr Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Tyr Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Asp Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Lys Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Gln Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Lys Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Tyr Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp Val Phe Glu 290 295 300 Lys Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asn Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Thr Asn Leu Asn Pro Arg Ile Ile Thr Pro Leu Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Phe Ser Lys Gly Ile Arg 420 <210> 49 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E10 subtype <400> 49 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Lys 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asn Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Thr Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Lys Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Asn Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Gln 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asn Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asn Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Thr Asn Leu Asn Pro Arg Ile Ile Lys Gln Leu Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Ser Lys Gly Ile Thr 420 <210> 50 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E11 subtype <400> 50 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Lys 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asn Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Thr Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Lys Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Thr Asp Leu Asn Ile Ile Thr Asn Ala Gln Ser Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asp Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Gln 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Val Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asp Ala Asn Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Lys Thr Tyr Ile 340 345 350 Gly His His Lys Tyr Phe Arg Leu Ser Asp Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Asp Gly Tyr Asn Ile Asn Thr Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Thr Asn Leu Asn Thr Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Val Val Arg Lys Ile Ile Arg Phe Cys Thr Asn Ile Phe 405 410 415 Ser Pro Lys Gly Ile Arg 420 <210> 51 <211> 425 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E1 subtype <400> 51 Met Leu Tyr Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val 1 5 10 15 Asn Asp Arg Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe 20 25 30 Tyr Lys Ser Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg 35 40 45 Asn Val Ile Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu 50 55 60 Lys Asn Gly Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp 65 70 75 80 Glu Glu Lys Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg 85 90 95 Ile Asn Asn Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys 100 105 110 Ala Asn Pro Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His 115 120 125 Ile Gly Asp Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln 130 135 140 Asp Ile Leu Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu 145 150 155 160 Phe Glu Thr Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro 165 170 175 Ser Asn His Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu 180 185 190 Tyr Ser Phe Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp 195 200 205 Pro Ala Leu Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu 210 215 220 Tyr Gly Ala Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Lys Gln 225 230 235 240 Asn Pro Leu Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu 245 250 255 Thr Phe Gly Gly Thr Asp Leu Asn Ile Ile Thr Ser Ala Gln Ser Asn 260 265 270 Asp Ile Tyr Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys 275 280 285 Leu Ser Lys Val Gln Val Ser Asn Pro Leu Leu Asn Pro Tyr Lys Asp 290 295 300 Val Phe Glu Ala Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr 305 310 315 320 Ser Val Asn Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser 325 330 335 Phe Thr Glu Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln 340 345 350 Thr Tyr Ile Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn 355 360 365 Asp Ser Ile Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys 370 375 380 Val Asn Phe Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr 385 390 395 400 Pro Ile Thr Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys 405 410 415 Asn Ile Val Ser Val Lys Gly Ile Arg 420 425 <210> 52 <211> 425 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E10 subtype <400> 52 Met Leu Tyr Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val 1 5 10 15 Asn Asp Lys Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe 20 25 30 Tyr Lys Ser Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg 35 40 45 Asn Val Ile Gly Thr Ile Pro Gln Asp Phe Leu Pro Pro Thr Ser Leu 50 55 60 Lys Asn Gly Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn 65 70 75 80 Glu Glu Lys Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg 85 90 95 Ile Asn Asp Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys 100 105 110 Ala Asn Pro Tyr Leu Gly Asn Asp Asn Thr Pro Asn Asn Gln Phe His 115 120 125 Ile Gly Asp Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln 130 135 140 Ser Ile Leu Leu Pro Thr Val Ile Ile Met Gly Ala Glu Pro Asp Leu 145 150 155 160 Phe Glu Thr Asn Ser Ser Asn Ile Ser Leu Lys Asn Asn Tyr Met Pro 165 170 175 Ser Asn His Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu 180 185 190 Tyr Ser Phe Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp 195 200 205 Pro Ala Leu Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu 210 215 220 Tyr Gly Ala Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln 225 230 235 240 Asn Pro Leu Ile Thr Asn Ile Arg Gly Thr Asn Ile Glu Glu Phe Leu 245 250 255 Thr Phe Gly Gly Thr Asp Leu Asn Ile Ile Thr Asn Ala Gln Ser Asn 260 265 270 Asp Ile Tyr Thr Asn Leu Leu Ala Asp Tyr Lys Lys Ile Ala Ser Lys 275 280 285 Leu Ser Gln Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp 290 295 300 Ile Phe Gln Glu Lys Tyr Gly Leu Asp Lys Asn Ala Ser Gly Ile Tyr 305 310 315 320 Ser Val Asn Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser 325 330 335 Phe Thr Glu Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Gln 340 345 350 Thr Tyr Ile Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn 355 360 365 Asn Ser Ile Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Thr Leu Lys 370 375 380 Val Asn Phe Arg Gly Gln Asn Thr Asn Leu Asn Pro Arg Ile Ile Thr 385 390 395 400 Gln Leu Thr Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys 405 410 415 Asn Ile Val Phe Ser Lys Gly Ile Thr 420 425 <210> 53 <211> 425 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E-E3 subtype <400> 53 Met Leu Tyr Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val 1 5 10 15 Asn Asp Arg Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe 20 25 30 Tyr Lys Ser Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg 35 40 45 Asn Val Ile Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu 50 55 60 Lys Asn Gly Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp 65 70 75 80 Glu Glu Lys Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg 85 90 95 Ile Asn Asn Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys 100 105 110 Ala Asn Pro Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His 115 120 125 Ile Gly Asp Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln 130 135 140 His Ile Leu Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu 145 150 155 160 Phe Glu Thr Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro 165 170 175 Ser Asn His Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu 180 185 190 Tyr Ser Phe Arg Phe Asn Asp Asn Ser Ile Asn Glu Phe Ile Gln Asp 195 200 205 Pro Ala Leu Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu 210 215 220 Tyr Gly Ala Lys Gly Ile Thr Thr Thr Cys Ile Ile Thr Gln Gln Gln 225 230 235 240 Asn Pro Leu Ile Thr Asn Arg Lys Gly Ile Asn Ile Glu Glu Phe Leu 245 250 255 Thr Phe Gly Gly Asn Asp Leu Asn Ile Ile Thr Val Ala Gln Tyr Asn 260 265 270 Asp Ile Tyr Thr Asn Leu Leu Asn Asp Tyr Arg Lys Ile Ala Ser Lys 275 280 285 Leu Ser Lys Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp 290 295 300 Ile Phe Gln Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr 305 310 315 320 Ser Val Asn Ile Asn Lys Phe Asp Asp Ile Phe Lys Lys Leu Tyr Ser 325 330 335 Phe Thr Glu Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu 340 345 350 Thr Tyr Ile Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn 355 360 365 Asp Ser Ile Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys 370 375 380 Val Asn Phe Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Lys 385 390 395 400 Pro Ile Thr Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys 405 410 415 Asn Ile Val Ser Val Lys Gly Ile Arg 420 425 <210> 54 <211> 425 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 54 Met Leu Tyr Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val 1 5 10 15 Asn Asp Arg Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe 20 25 30 Tyr Lys Ser Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg 35 40 45 Asn Val Ile Gly Thr Ile Pro Gln Asp Phe Gln Pro Pro Thr Ser Leu 50 55 60 Lys Asn Gly Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn 65 70 75 80 Glu Glu Lys Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg 85 90 95 Ile Asn Asp Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys 100 105 110 Ala Asn Pro Tyr Leu Gly Asn Asp Asn Thr Pro Asp Gly Asp Phe Ile 115 120 125 Ile Asn Asp Ala Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser Gln 130 135 140 Ser Ile Leu Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu 145 150 155 160 Phe Glu Thr Asn Ser Ser Asn Ile Ser Leu Ile Asn Asn Tyr Arg Pro 165 170 175 Ser Asn His Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu 180 185 190 Tyr Ser Phe Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp 195 200 205 Pro Ala Leu Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu 210 215 220 Tyr Gly Ala Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln 225 230 235 240 Asn Ser Leu Ile Thr Asn Ile Arg Gly Ile Asn Ile Glu Glu Phe Leu 245 250 255 Thr Phe Gly Gly Asn Asp Leu Asn Ile Ile Thr Ser Ser Gln Phe Asn 260 265 270 Asp Ile Tyr Thr Asn Leu Leu Asp Asp Tyr Lys Lys Ile Ala Ser Lys 275 280 285 Leu Ser Gln Val Arg Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp 290 295 300 Val Phe Gln Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr 305 310 315 320 Ser Val Asn Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser 325 330 335 Phe Thr Glu Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu 340 345 350 Thr Tyr Ile Gly Gln Tyr Lys Tyr Phe Gln Leu Ser Asn Leu Leu Asn 355 360 365 Asp Ser Ile Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys 370 375 380 Val Asn Phe Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr 385 390 395 400 Pro Ile Thr Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys 405 410 415 Asn Ile Val Ser Val Lys Gly Ile Arg 420 425 <210> 55 <211> 421 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 55 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Pro Val Asn Asp Arg Thr 1 5 10 15 Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser Phe 20 25 30 Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile Gly 35 40 45 Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly Asp 50 55 60 Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys Asp 65 70 75 80 Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn Asn 85 90 95 Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro Tyr 100 105 110 Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp Ala 115 120 125 Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln His Ile Leu Leu 130 135 140 Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr Asn 145 150 155 160 Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His Gly 165 170 175 Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe Arg 180 185 190 Phe Asn Asp Asn Ser Ile Asn Glu Phe Ile Gln Asp Pro Ala Leu Thr 195 200 205 Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala Lys 210 215 220 Gly Ile Thr Thr Thr Cys Ile Ile Thr Gln Gln Gln Asn Pro Leu Ile 225 230 235 240 Thr Asn Arg Lys Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly Gly 245 250 255 Asn Asp Leu Asn Ile Ile Thr Val Ala Gln Tyr Asn Asp Ile Tyr Thr 260 265 270 Asn Leu Leu Asn Asp Tyr Arg Lys Ile Ala Ser Lys Leu Ser Lys Val 275 280 285 Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln Glu 290 295 300 Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn Ile 305 310 315 320 Asn Lys Phe Asp Asp Ile Leu Lys Lys Leu Tyr Ser Phe Thr Glu Phe 325 330 335 Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile Gly 340 345 350 Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile Tyr 355 360 365 Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe Arg 370 375 380 Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Lys Pro Ile Thr Gly 385 390 395 400 Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val Ser 405 410 415 Val Lys Gly Ile Arg 420 <210> 56 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 56 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Glu Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Thr Pro Gln Asp Phe His Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asp Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asn 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Asn Gln Phe His Ile Gly Asp 115 120 125 Ala Ser Ala Val Glu Ile Lys Phe Ser Asn Gly Ser Gln His Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Ile Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Thr Cys Ile Ile Thr Gln Gln Gln Asn Pro Leu 225 230 235 240 Ile Thr Asn Arg Lys Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Asn Asp Leu Asn Ile Ile Thr Val Ala Gln Tyr Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asn Asp Tyr Arg Lys Ile Ala Ser Lys Leu Ser Lys 275 280 285 Val Gln Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Ile Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe His Asp Ile Leu Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Lys Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Lys Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 57 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> BoNT/E <400> 57 Met Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp Arg 1 5 10 15 Thr Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gln Gln Phe Tyr Lys Ser 20 25 30 Phe Asn Ile Met Lys Asn Ile Trp Ile Ile Pro Glu Arg Asn Val Ile 35 40 45 Gly Thr Ile Pro Gln Asp Phe Gln Pro Pro Thr Ser Leu Lys Asn Gly 50 55 60 Asp Ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gln Ser Asn Glu Glu Lys 65 70 75 80 Asp Arg Phe Leu Lys Ile Val Thr Lys Ile Phe Asn Arg Ile Asn Asp 85 90 95 Asn Leu Ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro 100 105 110 Tyr Leu Gly Asn Asp Asn Thr Pro Asp Gly Asp Phe Ile Ile Asn Asp 115 120 125 Ala Ser Ala Val Pro Ile Gln Phe Ser Asn Gly Ser Gln Ser Ile Leu 130 135 140 Leu Pro Asn Val Ile Ile Met Gly Ala Glu Pro Asp Leu Phe Glu Thr 145 150 155 160 Asn Ser Ser Asn Ile Ser Leu Ile Asn Asn Tyr Arg Pro Ser Asn His 165 170 175 Gly Phe Gly Ser Ile Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe 180 185 190 Arg Phe Asn Asp Asn Ser Met Asn Glu Phe Ile Gln Asp Pro Ala Leu 195 200 205 Thr Leu Met His Glu Leu Ile His Ser Leu His Gly Leu Tyr Gly Ala 210 215 220 Lys Gly Ile Thr Thr Lys Tyr Thr Ile Thr Gln Gln Gln Asn Ser Leu 225 230 235 240 Ile Thr Asn Ile Arg Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly 245 250 255 Gly Asn Asp Leu Asn Ile Ile Thr Ser Ser Gln Phe Asn Asp Ile Tyr 260 265 270 Thr Asn Leu Leu Asp Asp Tyr Lys Lys Ile Ala Ser Lys Leu Ser Gln 275 280 285 Val Arg Val Ser Asn Pro Gln Leu Asn Pro Tyr Lys Asp Val Phe Gln 290 295 300 Glu Lys Tyr Gly Leu Asp Lys Asp Ala Ser Gly Ile Tyr Ser Val Asn 305 310 315 320 Ile Asn Lys Phe Asn Asp Ile Phe Lys Lys Leu Tyr Ser Phe Thr Glu 325 330 335 Phe Asp Leu Ala Thr Lys Phe Gln Val Lys Cys Arg Glu Thr Tyr Ile 340 345 350 Gly Gln Tyr Lys Tyr Phe Gln Leu Ser Asn Leu Leu Asn Asp Ser Ile 355 360 365 Tyr Asn Ile Ser Glu Gly Tyr Asn Ile Asn Asn Leu Lys Val Asn Phe 370 375 380 Arg Gly Gln Asn Ala Asn Leu Asn Pro Arg Ile Ile Thr Pro Ile Thr 385 390 395 400 Gly Arg Gly Leu Val Lys Lys Ile Ile Arg Phe Cys Lys Asn Ile Val 405 410 415 Ser Val Lys Gly Ile Arg 420 <210> 58 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 58 Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Lys Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asp Glu Phe 115 120 125 Ser Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Leu Ser Thr Asn 130 135 140 Val Glu Ser Ser Met Leu Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Glu Ser Cys Cys Tyr Pro Val Arg Lys Leu Ile Asp Pro 165 170 175 Asp Val Val Tyr Asp Pro Ser Asn Tyr Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Glu Glu Thr Ile Glu Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ser Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Glu Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Ser Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val 420 425 430 Ile Pro Arg Lys 435 <210> 59 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 59 Met Pro Val Val Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asp Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Glu Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Glu Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Glu His Thr Pro Ile Asn Glu Phe 115 120 125 His Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Ser Ser Thr Asn 130 135 140 Val Lys Ser Ser Ile Ile Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Glu Asn Ser Ser Tyr Pro Val Arg Lys Leu Met Asp Ser 165 170 175 Gly Gly Val Tyr Asp Pro Ser Asn Asp Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly Tyr Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Lys Glu Thr Ile Lys Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Arg Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ile Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Asn Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val 420 425 430 Ile Pro Arg Lys 435 <210> 60 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F-F6 subtype <400> 60 Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Lys Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asp Glu Phe 115 120 125 Ser Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Leu Ser Thr Asn 130 135 140 Val Glu Ser Ser Met Leu Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Glu Ser Cys Cys Tyr Pro Val Arg Lys Leu Ile Asp Pro 165 170 175 Asp Val Val Tyr Asp Pro Ser Asn Tyr Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Glu Glu Thr Ile Glu Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ser Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Ser Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val 420 425 430 Ile Pro Arg Lys 435 <210> 61 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 61 Met Pro Val Val Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Glu Thr Ile Leu Tyr Met Gln Lys Pro Tyr Glu Glu Arg Ser Arg Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Pro Asn Val Trp Ile Met Pro Glu 35 40 45 Arg Asp Thr Ile Gly Thr Lys Pro Asp Glu Phe Gln Val Pro Asp Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Met Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Asn Pro Thr Gly Lys Val Leu Leu Glu Glu Val Ser 100 105 110 Asn Ala Arg Pro Tyr Leu Gly Asp Asp Asp Thr Leu Ile Asn Glu Phe 115 120 125 Phe Pro Val Asn Val Thr Thr Ser Val Asn Ile Lys Phe Ser Thr Asp 130 135 140 Val Glu Ser Ser Ile Ile Ser Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Lys Ala Tyr Cys Thr Pro Leu Val Arg Phe Asn Lys Ser 165 170 175 Asp Lys Leu Ile Glu Pro Ser Asn His Gly Phe Gly Ser Ile Asn Ile 180 185 190 Leu Thr Phe Ser Pro Glu Tyr Glu His Ile Phe Asn Asp Ile Ser Gly 195 200 205 Gly Asn His Asn Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Lys 225 230 235 240 Ala Val Thr His Lys Glu Ser Leu Val Ala Glu Arg Gly Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Glu Asp Leu Asn Ile Ile Pro Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asp Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Arg Glu Val 290 295 300 Asn Thr Ala Pro Pro Gly Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Arg Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Arg Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ile Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Val Lys Val Pro Asp Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Asn Ile Asn Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Ile Lys Phe Cys Lys Ser Ile 420 425 430 Ile Pro Arg Lys 435 <210> 62 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F-F6 subtype <400> 62 Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Lys Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Lys Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asp Glu Phe 115 120 125 Ser Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Leu Ser Thr Asn 130 135 140 Val Glu Ser Ser Met Leu Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Glu Ser Cys Cys Tyr Pro Val Arg Lys Leu Ile Asp Pro 165 170 175 Asp Val Val Tyr Asp Pro Ser Asn Tyr Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Glu Glu Thr Ile Glu Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ser Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Ser Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val 420 425 430 Ile Pro Arg Lys 435 <210> 63 <211> 447 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F-F9 subtype <400> 63 Met Asp Ile Glu Arg Ser Ile Gly Gly Ile Phe Met Pro Phe Glu Ile 1 5 10 15 Asn Ser Phe Asn Tyr Asp Asp Pro Val Asn Asp Glu Thr Ile Leu Tyr 20 25 30 Met Gln Lys Pro Tyr Glu Gly Lys Ser Lys Lys Tyr Tyr Lys Ala Phe 35 40 45 Glu Ile Met Pro Asn Val Trp Ile Met Pro Glu Arg Asp Thr Ile Gly 50 55 60 Thr Glu Pro Ser Asp Phe Gln Val Pro Val Ser Leu Lys Asn Gly Ser 65 70 75 80 Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr Asp Ala Glu Lys Asp 85 90 95 Arg Tyr Leu Lys Ile Met Ile Lys Leu Phe Asn Arg Ile Asn Ser Asn 100 105 110 Pro Ala Gly Glu Val Leu Leu Arg Glu Ile Ser Lys Ala Arg Pro Tyr 115 120 125 Leu Gly Asp Glu His Thr Pro Ile Asn Glu Phe Phe Pro Val Asn Val 130 135 140 Thr Thr Ser Val Asn Ile Lys Phe Ser Asp Asp Val Lys Ser Ser Ile 145 150 155 160 Ile Leu Asn Leu Leu Val Leu Gly Ala Gly Pro Asp Ile Phe Lys Val 165 170 175 Phe Cys Ser Ser Leu Ala Arg Ser Ile Asp Ser Gly Glu Phe Tyr Gln 180 185 190 Pro Ser Asn His Gly Phe Gly Ser Ile Asn Ile Leu Thr Phe Ser Pro 195 200 205 Glu Tyr Glu His Ile Phe Asn Asp Ile Ser Gly Gly Asp His Asn Ser 210 215 220 Ile Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser Leu Ala His Glu Leu 225 230 235 240 Ile His Ala Leu His Gly Leu Tyr Gly Ala Lys Ala Val Thr Tyr Lys 245 250 255 Glu Thr Ile Glu Ala Lys Arg Gly Pro Leu Met Ile Ala Glu Lys Pro 260 265 270 Ile Arg Val Glu Glu Phe Leu Thr Phe Gly Gly Lys Asp Leu Asn Ile 275 280 285 Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn Asn Leu Leu Asp Asn 290 295 300 Tyr Glu Lys Ile Ala Thr Arg Leu Ser Asn Val Asn Thr Ala Pro Ser 305 310 315 320 Gly Tyr Asp Ile Asn Lys Tyr Lys Asp Ile Phe Gln Trp Lys Tyr Gly 325 330 335 Leu Asp Glu Asn Ala Asp Gly Ser Tyr Thr Val Asn Lys Asn Lys Phe 340 345 350 Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr Glu Ile Asp Leu Ala 355 360 365 Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr Phe Ile Glu Tyr Gly 370 375 380 Phe Val Lys Val Pro Asp Leu Leu Asp Asp Asp Ile Tyr Thr Val Ser 385 390 395 400 Glu Gly Phe Asn Ile Asp Asn Leu Ala Val Asn Asn Arg Gly Gln Asn 405 410 415 Ile Asn Leu Asn Pro Lys Ile Ile Gly Ser Ile Pro Asp Glu Gly Leu 420 425 430 Val Glu Lys Ile Val Lys Phe Cys Lys Asn Ile Leu Pro Arg Lys 435 440 445 <210> 64 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 64 Met Pro Val Val Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Glu Thr Ile Leu Tyr Met Gln Lys Pro Tyr Glu Glu Arg Ser Arg Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Pro Asn Val Trp Ile Met Pro Glu 35 40 45 Arg Asp Thr Ile Gly Thr Lys Pro Asp Asp Phe Gln Val Pro Asp Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Met Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Asn Pro Thr Gly Lys Val Leu Leu Glu Glu Val Ser 100 105 110 Asn Ala Arg Pro Tyr Leu Gly Asp Asp Asp Thr Leu Ile Asn Glu Phe 115 120 125 Phe Pro Val Asn Val Thr Thr Ser Val Asn Ile Lys Phe Ser Thr Asp 130 135 140 Val Glu Ser Ser Ile Ile Ser Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Lys Ala Tyr Cys Thr Pro Leu Val Arg Phe Asn Lys Ser 165 170 175 Asp Lys Leu Ile Glu Pro Ser Asn His Gly Phe Gly Ser Ile Asn Ile 180 185 190 Leu Thr Phe Ser Pro Glu Tyr Glu His Ile Phe Asn Asp Ile Ser Gly 195 200 205 Gly Asp His Asn Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Lys 225 230 235 240 Ala Val Thr His Lys Glu Thr Ile Glu Val Lys Arg Gly Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Glu Asp Leu Asn Ile Ile Pro Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asp Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Arg Glu Val 290 295 300 Asn Thr Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Arg Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Arg Asn Lys Phe Asn Gly Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ile Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Val Lys Val Pro Asp Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Asn Ile Asn Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Ile Lys Phe Cys Lys Ser Ile 420 425 430 Ile Pro Arg Lys 435 <210> 65 <211> 428 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F-F7 subtype <400> 65 Met Pro Val Asn Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asn Asn 1 5 10 15 Thr Thr Ile Leu Tyr Met Lys Met Pro Tyr Tyr Glu Asp Ser Asn Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Asp Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Ile Ile Gly Lys Lys Pro Ser Asp Phe Tyr Pro Pro Ile Ser 50 55 60 Leu Asp Ser Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Phe Leu Lys Thr Val Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Gln Val Leu Leu Glu Glu Ile Lys 100 105 110 Asn Gly Lys Pro Tyr Leu Gly Asn Asp His Thr Ala Val Asn Glu Phe 115 120 125 Cys Ala Asn Asn Arg Ser Thr Ser Val Glu Ile Lys Glu Ser Asn Gly 130 135 140 Thr Thr Asp Ser Met Leu Leu Asn Leu Val Ile Leu Gly Pro Gly Pro 145 150 155 160 Asn Ile Leu Glu Cys Ser Thr Phe Pro Val Arg Ile Phe Pro Asn Asn 165 170 175 Ile Ala Tyr Asp Pro Ser Glu Lys Gly Phe Gly Ser Ile Gln Leu Met 180 185 190 Ser Phe Ser Thr Glu Tyr Glu Tyr Ala Phe Asn Asp Asn Thr Asp Leu 195 200 205 Phe Ile Ala Asp Pro Ala Ile Ser Leu Ala His Glu Leu Ile His Val 210 215 220 Leu His Gly Leu Tyr Gly Ala Lys Gly Val Thr Asn Lys Lys Val Ile 225 230 235 240 Glu Val Asp Gln Gly Ala Leu Met Ala Ala Glu Lys Asp Ile Lys Ile 245 250 255 Glu Glu Phe Ile Thr Phe Gly Gly Gln Asp Leu Asn Ile Ile Thr Asn 260 265 270 Ser Thr Asn Gln Lys Ile Tyr Asp Asn Leu Leu Ser Asn Tyr Thr Ala 275 280 285 Ile Ala Ser Arg Leu Ser Gln Val Asn Ile Asn Asn Ser Ala Leu Asn 290 295 300 Thr Thr Tyr Tyr Lys Asn Phe Phe Gln Trp Lys Tyr Gly Leu Asp Gln 305 310 315 320 Asp Ser Asn Gly Asn Tyr Thr Val Asn Ile Ser Lys Phe Asn Ala Ile 325 330 335 Tyr Lys Lys Leu Phe Ser Phe Thr Glu Cys Asp Leu Ala Gln Lys Phe 340 345 350 Gln Val Lys Asn Arg Ser Asn Tyr Leu Phe His Phe Lys Pro Phe Lys 355 360 365 Leu Leu Asp Leu Leu Asp Asp Asn Ile Tyr Ser Ile Ser Glu Gly Phe 370 375 380 Asn Ile Gly Ser Leu Arg Val Asn Asn Asn Gly Gln Asn Ile Asn Leu 385 390 395 400 Asn Ser Arg Ile Val Gly Pro Ile Pro Asp Asn Gly Leu Val Glu Arg 405 410 415 Phe Val Gly Leu Cys Lys Ser Ile Val Ser Lys Lys 420 425 <210> 66 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 66 Met Pro Val Val Ile Asn Ser Phe Asn Tyr Asp Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Met Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asn Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Lys Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Glu Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Asp His Thr Pro Ile Asn Glu Phe 115 120 125 His Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Ser Ser Thr Asn 130 135 140 Val Glu Ser Ser Ile Ile Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asn Ile Phe Glu Asn Ser Ser Tyr Pro Val Arg Lys Leu Met Asn Ser 165 170 175 Gly Glu Val Tyr Asp Pro Ser Asn Asp Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly His Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Lys Glu Thr Ile Lys Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asp Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Glu Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asn Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ile Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Asn Ile Asn Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Leu Cys Lys Ser Ile 420 425 430 Ile Pro Arg Lys 435 <210> 67 <211> 428 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 67 Met Pro Val Asn Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asn Asn 1 5 10 15 Thr Thr Ile Leu Tyr Met Lys Met Pro Tyr Tyr Glu Asp Ser Asn Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Asp Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Ile Ile Gly Lys Lys Pro Ser Asp Phe Tyr Pro Pro Ile Ser 50 55 60 Leu Asp Ser Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Phe Leu Lys Thr Val Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Gln Val Leu Leu Glu Glu Ile Lys 100 105 110 Asn Gly Lys Pro Tyr Leu Gly Asn Asp His Thr Ala Val Asn Glu Phe 115 120 125 Cys Ala Asn Asn Arg Ser Thr Ser Val Glu Ile Lys Glu Ser Lys Gly 130 135 140 Thr Thr Asp Ser Met Leu Leu Asn Leu Val Ile Leu Gly Pro Gly Pro 145 150 155 160 Asn Ile Leu Glu Cys Ser Thr Phe Pro Val Arg Ile Phe Pro Asn Asn 165 170 175 Ile Ala Tyr Asp Pro Ser Glu Lys Gly Phe Gly Ser Ile Gln Leu Met 180 185 190 Ser Phe Ser Thr Glu Tyr Glu Tyr Ala Phe Asn Asp Asn Thr Asp Leu 195 200 205 Phe Ile Ala Asp Pro Ala Ile Ser Leu Ala His Glu Leu Ile His Val 210 215 220 Leu His Gly Leu Tyr Gly Ala Lys Gly Val Thr Asn Lys Lys Val Ile 225 230 235 240 Glu Val Asp Gln Gly Ala Leu Met Ala Ala Glu Lys Asp Ile Lys Ile 245 250 255 Glu Glu Phe Ile Thr Phe Gly Gly Gln Asp Leu Asn Ile Ile Thr Asn 260 265 270 Ser Thr Asn Gln Lys Ile Tyr Asp Asn Leu Leu Ser Asn Tyr Thr Ala 275 280 285 Ile Ala Ser Arg Leu Ser Gln Val Asn Ile Asn Asn Ser Ala Leu Asn 290 295 300 Thr Thr Tyr Tyr Lys Asn Phe Phe Gln Trp Lys Tyr Gly Leu Asp Gln 305 310 315 320 Asp Ser Asn Gly Asn Tyr Thr Val Asn Ile Ser Lys Phe Asn Ala Ile 325 330 335 Tyr Lys Lys Leu Phe Ser Phe Thr Glu Cys Asp Leu Ala Gln Lys Phe 340 345 350 Gln Val Lys Asn Arg Ser Asn Tyr Leu Phe His Phe Lys Pro Phe Arg 355 360 365 Leu Leu Asp Leu Leu Asp Asp Asn Ile Tyr Ser Ile Ser Glu Gly Phe 370 375 380 Asn Ile Gly Ser Leu Arg Val Asn Asn Asn Gly Gln Asn Ile Asn Leu 385 390 395 400 Asn Ser Arg Ile Val Gly Pro Ile Pro Asp Asn Gly Leu Val Glu Arg 405 410 415 Phe Val Gly Leu Cys Lys Ser Ile Val Ser Lys Lys 420 425 <210> 68 <211> 428 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 68 Met Pro Val Asn Ile Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asn Asn 1 5 10 15 Thr Thr Ile Leu Tyr Met Lys Met Pro Tyr Tyr Glu Asp Ser Asn Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Asp Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Ile Ile Gly Lys Lys Pro Ser Asp Phe Tyr Pro Pro Ile Ser 50 55 60 Leu Asp Ser Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Phe Leu Lys Thr Val Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Gln Val Leu Leu Glu Glu Ile Lys 100 105 110 Asn Gly Lys Pro Tyr Leu Gly Asn Asp His Thr Ala Val Asn Glu Phe 115 120 125 Cys Ala Asn Asn Arg Ser Thr Ser Val Glu Ile Lys Glu Ser Asn Gly 130 135 140 Thr Thr Asp Ser Met Leu Leu Asn Leu Val Ile Leu Gly Pro Gly Pro 145 150 155 160 Asn Ile Leu Glu Cys Ser Thr Phe Pro Val Arg Ile Phe Pro Asn Asn 165 170 175 Ile Ala Tyr Asp Pro Ser Glu Lys Gly Phe Gly Ser Ile Gln Leu Met 180 185 190 Ser Phe Ser Thr Glu Tyr Glu Tyr Ala Phe Asn Asp Asn Thr Asp Leu 195 200 205 Phe Ile Ala Asp Pro Ala Ile Ser Leu Ala His Glu Leu Ile His Val 210 215 220 Leu His Gly Leu Tyr Gly Ala Lys Gly Val Thr Asn Lys Lys Val Ile 225 230 235 240 Glu Val Asp Gln Gly Ala Leu Met Ala Ala Glu Lys Asp Ile Lys Ile 245 250 255 Glu Glu Phe Ile Thr Phe Gly Gly Gin Asp Leu Asn Ile Val Thr Asn 260 265 270 Ser Thr Asn Gln Lys Ile Tyr Asp Asn Leu Leu Ser Asn Tyr Thr Ala 275 280 285 Ile Ala Ser Arg Leu Ser Gln Val Asn Ile Asn Asn Ser Ala Leu Asn 290 295 300 Thr Thr Tyr Tyr Lys Asn Phe Phe Gln Trp Lys Tyr Gly Leu Asp Gln 305 310 315 320 Asp Ser Asn Gly Asn Tyr Thr Val Asn Ile Ser Lys Phe Asn Ala Ile 325 330 335 Tyr Lys Lys Leu Phe Ser Phe Thr Glu Cys Asp Leu Ala Gln Lys Phe 340 345 350 Gln Val Lys Asn Arg Ser Asn Tyr Leu Phe His Phe Lys Pro Phe Lys 355 360 365 Leu Leu Asp Leu Leu Asp Asp Asn Ile Tyr Ser Ile Ser Glu Gly Phe 370 375 380 Asn Ile Gly Ser Leu Arg Val Asn Asn Asn Gly Gln Asn Ile Asn Leu 385 390 395 400 Asn Ser Arg Ile Val Gly Pro Ile Pro Asp Asn Gly Leu Val Glu Arg 405 410 415 Phe Val Gly Leu Cys Lys Ser Ile Val Ser Lys Lys 420 425 <210> 69 <211> 436 <212> PRT <213> Artificial Sequence <220> <223> BoNT/F <400> 69 Met Pro Val Ala Ile Asn Ser Phe Asn Tyr Asn Asp Pro Val Asn Asp 1 5 10 15 Asp Thr Ile Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys 20 25 30 Tyr Tyr Lys Ala Phe Glu Ile Met Arg Asn Val Trp Ile Ile Pro Glu 35 40 45 Arg Asn Thr Ile Gly Thr Asp Pro Ser Asp Phe Asp Pro Pro Ala Ser 50 55 60 Leu Glu Asn Gly Ser Ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr 65 70 75 80 Asp Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr Ile Lys Leu Phe Lys 85 90 95 Arg Ile Asn Ser Asn Pro Ala Gly Glu Val Leu Leu Gln Glu Ile Ser 100 105 110 Tyr Ala Lys Pro Tyr Leu Gly Asn Glu His Thr Pro Ile Asn Glu Phe 115 120 125 His Pro Val Thr Arg Thr Thr Ser Val Asn Ile Lys Ser Ser Thr Asn 130 135 140 Val Lys Ser Ser Ile Ile Leu Asn Leu Leu Val Leu Gly Ala Gly Pro 145 150 155 160 Asp Ile Phe Glu Asn Ser Ser Tyr Pro Val Arg Lys Leu Met Asp Ser 165 170 175 Gly Gly Val Tyr Asp Pro Ser Asn Asp Gly Phe Gly Ser Ile Asn Ile 180 185 190 Val Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp Ile Ser Gly 195 200 205 Gly Tyr Asn Ser Ser Thr Glu Ser Phe Ile Ala Asp Pro Ala Ile Ser 210 215 220 Leu Ala His Glu Leu Ile His Ala Leu His Gly Leu Tyr Gly Ala Arg 225 230 235 240 Gly Val Thr Tyr Lys Glu Thr Ile Lys Val Lys Gln Ala Pro Leu Met 245 250 255 Ile Ala Glu Lys Pro Ile Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly 260 265 270 Gln Asp Leu Asn Ile Ile Thr Ser Ala Met Lys Glu Lys Ile Tyr Asn 275 280 285 Asn Leu Leu Ala Asn Tyr Glu Lys Ile Ala Thr Arg Leu Ser Arg Val 290 295 300 Asn Ser Ala Pro Pro Glu Tyr Asp Ile Asn Glu Tyr Lys Asp Tyr Phe 305 310 315 320 Gln Trp Lys Tyr Gly Leu Asp Lys Asn Ala Asp Gly Ser Tyr Thr Val 325 330 335 Asn Glu Asn Lys Phe Asn Glu Ile Tyr Lys Lys Leu Tyr Ser Phe Thr 340 345 350 Glu Ile Asp Leu Ala Asn Lys Phe Lys Val Lys Cys Arg Asn Thr Tyr 355 360 365 Phe Ile Lys Tyr Gly Phe Leu Lys Val Pro Asn Leu Leu Asp Asp Asp 370 375 380 Ile Tyr Thr Val Ser Glu Gly Phe Asn Ile Gly Asn Leu Ala Val Asn 385 390 395 400 Asn Arg Gly Gln Asn Ile Lys Leu Asn Pro Lys Ile Ile Asp Ser Ile 405 410 415 Pro Asp Lys Gly Leu Val Glu Lys Ile Val Lys Phe Cys Lys Ser Val 420 425 430 Ile Pro Arg Lys 435 <210> 70 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> BoNT/G <400> 70 Met Pro Val Asn Ile Lys Phe Asn Tyr Asn Asp Pro Ile Asn Asn Asp 1 5 10 15 Asp Ile Ile Met Met Glu Pro Phe Asn Asp Pro Gly Pro Gly Thr Tyr 20 25 30 Tyr Lys Ala Phe Arg Ile Ile Asp Arg Ile Trp Ile Val Pro Glu Arg 35 40 45 Phe Thr Tyr Gly Phe Gln Pro Asp Gln Phe Asn Ala Ser Thr Gly Val 50 55 60 Phe Ser Lys Asp Val Tyr Glu Tyr Tyr Asp Pro Thr Tyr Leu Lys Thr 65 70 75 80 Asp Ala Glu Lys Asp Lys Phe Leu Lys Thr Met Ile Lys Leu Phe Asn 85 90 95 Arg Ile Asn Ser Lys Pro Ser Gly Gln Arg Leu Leu Asp Met Ile Val 100 105 110 Asp Ala Ile Pro Tyr Leu Gly Asn Ala Ser Thr Pro Pro Asp Lys Phe 115 120 125 Ala Ala Asn Val Ala Asn Val Ser Ile Asn Lys Lys Ile Ile Gln Pro 130 135 140 Gly Ala Glu Asp Gln Ile Lys Gly Leu Met Thr Asn Leu Ile Ile Phe 145 150 155 160 Gly Pro Gly Pro Val Leu Ser Asp Asn Phe Thr Asp Ser Met Ile Met 165 170 175 Asn Gly His Ser Pro Ile Ser Glu Gly Phe Gly Ala Arg Met Met Ile 180 185 190 Arg Phe Cys Pro Ser Cys Leu Asn Val Phe Asn Asn Val Gln Glu Asn 195 200 205 Lys Asp Thr Ser Ile Phe Ser Arg Arg Ala Tyr Phe Ala Asp Pro Ala 210 215 220 Leu Thr Leu Met His Glu Leu Ile His Val Leu His Gly Leu Tyr Gly 225 230 235 240 Ile Lys Ile Ser Asn Leu Pro Ile Thr Pro Asn Thr Lys Glu Phe Phe 245 250 255 Met Gln His Ser Asp Pro Val Gln Ala Glu Glu Leu Tyr Thr Phe Gly 260 265 270 Gly His Asp Pro Ser Val Ile Ser Pro Ser Thr Asp Met Asn Ile Tyr 275 280 285 Asn Lys Ala Leu Gln Asn Phe Gln Asp Ile Ala Asn Arg Leu Asn Ile 290 295 300 Val Ser Ser Ala Gln Gly Ser Gly Ile Asp Ile Ser Leu Tyr Lys Gln 305 310 315 320 Ile Tyr Lys Asn Lys Tyr Asp Phe Val Glu Asp Pro Asn Gly Lys Tyr 325 330 335 Ser Val Asp Lys Asp Lys Phe Asp Lys Leu Tyr Lys Ala Leu Met Phe 340 345 350 Gly Phe Thr Glu Thr Asn Leu Ala Gly Glu Tyr Gly Ile Lys Thr Arg 355 360 365 Tyr Ser Tyr Phe Ser Glu Tyr Leu Pro Pro Ile Lys Thr Glu Lys Leu 370 375 380 Leu Asp Asn Thr Ile Tyr Thr Gln Asn Glu Gly Phe Asn Ile Ala Ser 385 390 395 400 Lys Asn Leu Lys Thr Glu Phe Asn Gly Gln Asn Lys Ala Val Asn Lys 405 410 415 Glu Ala Tyr Glu Glu Ile Ser Leu Glu His Leu Val Ile Tyr Arg Ile 420 425 430 Ala Met Cys Lys Pro Val Met Tyr Lys 435 440 <210> 71 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> EGFP <400> 71 Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 1 5 10 15 Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly 20 25 30 Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile 35 40 45 Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 50 55 60 Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys 65 70 75 80 Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu 85 90 95 Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu 100 105 110 Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly 115 120 125 Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 130 135 140 Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn 145 150 155 160 Gly Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser 165 170 175 Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly 180 185 190 Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu 195 200 205 Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe 210 215 220 Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys 225 230 235 <210> 72 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> EYFP <400> 72 Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 1 5 10 15 Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly 20 25 30 Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile 35 40 45 Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 50 55 60 Phe Gly Tyr Gly Leu Gln Cys Phe Ala Arg Tyr Pro Asp His Met Lys 65 70 75 80 Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu 85 90 95 Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu 100 105 110 Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly 115 120 125 Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 130 135 140 Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn 145 150 155 160 Gly Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser 165 170 175 Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly 180 185 190 Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Tyr Gln Ser Ala Leu 195 200 205 Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe 210 215 220 Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys 225 230 235 <210> 73 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> ECFP <400> 73 Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 1 5 10 15 Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly 20 25 30 Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile 35 40 45 Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 50 55 60 Leu Thr Trp Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys 65 70 75 80 Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu 85 90 95 Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu 100 105 110 Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly 115 120 125 Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 130 135 140 Asn Tyr Ile Ser His Asn Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn 145 150 155 160 Gly Ile Lys Ala Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser 165 170 175 Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly 180 185 190 Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu 195 200 205 Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe 210 215 220 Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys 225 230 235 <210> 74 <211> 236 <212> PRT <213> Artificial Sequence <220> <223> ERFP <400> 74 Met Ser Cys Ser Lys Asn Val Ile Lys Glu Phe Met Arg Phe Lys Val 1 5 10 15 Arg Met Glu Gly Thr Val Asn Gly His Glu Phe Glu Ile Glu Gly Glu 20 25 30 Gly Glu Gly Arg Pro Tyr Glu Gly His Asn Thr Val Lys Leu Lys Val 35 40 45 Thr Lys Gly Gly Pro Leu Pro Phe Ala Trp Asp Ile Leu Ser Pro Gln 50 55 60 Phe Gln Tyr Gly Ser Lys Val Tyr Val Lys His Pro Ala Asp Ile Pro 65 70 75 80 Asp Tyr Lys Lys Leu Ser Phe Pro Glu Gly Phe Lys Trp Glu Arg Val 85 90 95 Met Asn Phe Glu Asp Gly Gly Val Val Thr Val Thr Gln Asp Ser Ser 100 105 110 Leu Gln Asp Gly Cys Phe Ile Tyr Lys Val Lys Phe Ile Gly Val Asn 115 120 125 Phe Pro Ser Asp Gly Pro Val Met Gln Lys Lys Thr Met Gly Trp Glu 130 135 140 Ala Ser Thr Glu Arg Leu Tyr Pro Arg Asp Gly Val Leu Lys Gly Glu 145 150 155 160 Ile His Lys Ala Leu Lys Leu Lys Asp Gly Gly His Tyr Leu Val Glu 165 170 175 Phe Lys Thr Ile Tyr Met Ala Lys Lys Pro Val Gln Leu Pro Gly Tyr 180 185 190 Tyr Tyr Val Asp Ser Lys Leu Asp Ile Thr Ser His Asn Lys Asp Tyr 195 200 205 Thr Ile Val Glu Gln Tyr Glu Arg Thr Glu Gly Arg His His Leu Phe 210 215 220 Leu Lys Ala Glu Leu Gly Ser Asn Val Gly Glu Arg 225 230 235 <210> 75 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> EBFP <400> 75 Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 1 5 10 15 Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly 20 25 30 Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile 35 40 45 Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 50 55 60 Leu Thr His Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys 65 70 75 80 Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu 85 90 95 Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu 100 105 110 Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly 115 120 125 Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 130 135 140 Asn Phe Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn 145 150 155 160 Gly Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser 165 170 175 Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly 180 185 190 Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu 195 200 205 Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe 210 215 220 Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys 225 230 235 <210> 76 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> His10 <400> 76 His His His His His His His His His His 1 5 10 <210> 77 <211> 26 <212> PRT <213> Artificial Sequence <220> <223> LAH4 <400> 77 Lys Lys Ala Leu Leu Ala Leu Ala Leu His His Leu Ala His Leu Ala 1 5 10 15 Leu His Leu Ala Leu Ala Leu Lys Lys Ala 20 25 <210> 78 <211> 26 <212> PRT <213> Artificial Sequence <220> <223> LAH4-L1 <400> 78 Lys Lys Ala Leu Leu Ala His Ala Leu His Leu Leu Ala Leu Leu Ala 1 5 10 15 Leu His Leu Ala His Ala Leu Lys Lys Ala 20 25 <210> 79 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> K6p <400> 79 Cys His Lys Lys Lys Lys Lys Lys Lys His Cys 1 5 10 <210> 80 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> H5WYG <400> 80 Gly Leu Phe His Ala Ile Ala His Phe Ile His Gly Gly Trp His Gly 1 5 10 15 Leu Ile His Gly Trp Tyr Gly 20 <210> 81 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> diINF-7 <400> 81 Gly Leu Phe His Ala Ile Ala His Phe Ile His Gly Gly Trp His Gly 1 5 10 15 Leu Ile His Gly Trp Tyr Gly 20 <210> 82 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> TAT <400> 82 Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Arg Arg Gln 1 5 10 <210> 83 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Glycoprotein <400> 83 Leu Ala Ser Thr Leu Thr Arg Trp Ala His Tyr Asn Ala Leu Ile Arg 1 5 10 15 Ala Phe <210> 84 <211> 26 <212> PRT <213> Artificial Sequence <220> <223> Melittin <400> 84 Gly Ile Gly Ala Val Leu Lys Val Leu Thr Thr Gly Leu Pro Ala Leu 1 5 10 15 Ile Ser Trp Ile Lys Arg Lys Arg Gln Gln 20 25 <210> 85 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> MT20 <400> 85 Gly Ile Gly Ala Val Leu Lys Val Leu Thr Thr Gly Leu Pro Ala Leu 1 5 10 15 Ile Ser Trp Ile 20 <210> 86 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Penetratin <400> 86 Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys 1 5 10 15 <210> 87 <211> 22 <212> PRT <213> Artificial Sequence <220> <223> R6-Penetratin <400> 87 Arg Arg Arg Arg Arg Arg Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg 1 5 10 15 Arg Met Lys Trp Lys Lys 20 <210> 88 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> <400> 88 Gly Trp Thr Leu Asn Ser Ala Gly Tyr Leu Leu Gly Lys Ile Asn Leu 1 5 10 15 Lys Ala Leu Ala Ala Leu Ala Lys Lys Ile Leu 20 25 <210> 89 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> TP10 <400> 89 Ala Gly Tyr Leu Leu Gly Lys Ile Asn Leu Lys Ala Leu Ala Ala Leu 1 5 10 15 Ala Lys Lys Ile Leu 20 <210> 90 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> EB1 <400> 90 Leu Ile Arg Leu Trp Ser His Leu Ile His Ile Trp Phe Gln Asn Arg 1 5 10 15 Arg Leu Lys Trp Lys Lys Lys 20 <210> 91 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Bovine prion protein <400> 91 Met Val Lys Ser Lys Ile Gly Ser Trp Ile Leu Val Leu Phe Val Ala 1 5 10 15 Met Trp Ser Asp Val Gly Leu Cys Lys Lys Arg Pro Lys Pro 20 25 30 <210> 92 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Aurein 1.2 <400> 92 Gly Leu Phe Asp Ile Ile Lys Lys Ile Ala Glu Ser Phe 1 5 10 <210> 93 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Proline-rich protein <400> 93 Cys Phe Val Arg Leu Pro Pro Pro Val Arg Leu Pro Pro Pro Val Arg 1 5 10 15 Leu Pro Pro Pro 20 <210> 94 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> GALA <400> 94 Trp Glu Ala Ala Leu Ala Glu Ala Leu Ala Glu Ala Leu Ala Glu His 1 5 10 15 Leu Ala Glu Ala Leu Ala Glu Ala Leu Glu Ala Leu Ala Ala 20 25 30 <210> 95 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> KALA <400> 95 Trp Glu Ala Lys Leu Ala Lys Ala Leu Ala Lys Ala Leu Ala Lys His 1 5 10 15 Leu Ala Lys Ala Leu Ala Lys Ala Leu Lys Ala Cys Glu Ala 20 25 30 <210> 96 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> JTS-1 <400> 96 Gly Leu Phe Glu Ala Leu Leu Glu Leu Leu Glu Ser Leu Trp Glu Leu 1 5 10 15 Leu Leu Glu Ala 20 <210> 97 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> ppTG1 <400> 97 Gly Leu Phe Lys Ala Leu Leu Lys Leu Leu Lys Ser Leu Trp Lys Leu 1 5 10 15 Leu Leu Lys Ala 20 <210> 98 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> ppTG20 <400> 98 Gly Leu Arg Lys Ala Leu Leu Arg Leu Leu Arg Ser Leu Trp Arg Leu 1 5 10 15 Leu Leu Arg Ala 20 <210> 99 <211> 427 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype RBD <400> 99 Met Lys Asn Ile Ile Asn Thr Ser Ile Leu Asn Leu Arg Tyr Glu Ser 1 5 10 15 Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile Gly 20 25 30 Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu Phe 35 40 45 Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile Val 50 55 60 Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg Ile 65 70 75 80 Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile Ile 85 90 95 Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr Gly 100 105 110 Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg Val 115 120 125 Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn Arg 130 135 140 Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys Ile 145 150 155 160 Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu Gly 165 170 175 Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys Arg 180 185 190 Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp Lys 195 200 205 Glu Leu Asn Glu Lys Glu Ile Lys Asp Leu Tyr Asp Asn Gln Ser Asn 210 215 220 Ser Gly Ile Leu Lys Asp Phe Trp Gly Asp Tyr Leu Gln Tyr Asp Lys 225 230 235 240 Pro Tyr Tyr Met Leu Asn Leu Tyr Asp Pro Asn Lys Tyr Val Asp Val 245 250 255 Asn Asn Val Gly Ile Arg Gly Tyr Met Tyr Leu Lys Gly Pro Arg Gly 260 265 270 Ser Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser Leu Tyr Arg Gly 275 280 285 Thr Lys Phe Ile Ile Lys Lys Tyr Ala Ser Gly Asn Lys Asp Asn Ile 290 295 300 Val Arg Asn Asn Asp Arg Val Tyr Ile Asn Val Val Val Lys Asn Lys 305 310 315 320 Glu Tyr Arg Leu Ala Thr Asn Ala Ser Gln Ala Gly Val Glu Lys Ile 325 330 335 Leu Ser Ala Leu Glu Ile Pro Asp Val Gly Asn Leu Ser Gln Val Val 340 345 350 Val Met Lys Ser Lys Asn Asp Gln Gly Ile Thr Asn Lys Cys Lys Met 355 360 365 Asn Leu Gln Asp Asn Asn Gly Asn Asp Ile Gly Phe Ile Gly Phe His 370 375 380 Gln Phe Asn Asn Ile Ala Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg 385 390 395 400 Gln Ile Glu Arg Ser Ser Arg Thr Leu Gly Cys Ser Trp Glu Phe Ile 405 410 415 Pro Val Asp Asp Gly Trp Gly Glu Arg Pro Leu 420 425 <210> 100 <211> 427 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype RBD <400> 100 Met Lys Asn Ile Val Asn Thr Ser Ile Leu Ser Ile Val Tyr Lys Lys 1 5 10 15 Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile Gly 20 25 30 Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu Ile 35 40 45 Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile Val 50 55 60 Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys Ile 65 70 75 80 Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile Ile 85 90 95 Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr Gly 100 105 110 Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg Val 115 120 125 Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn Arg 130 135 140 Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys Ile 145 150 155 160 Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu Gly 165 170 175 Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys Arg 180 185 190 Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp Lys 195 200 205 Glu Leu Asn Glu Lys Glu Ile Lys Asp Leu Tyr Asp Ser Gln Ser Asn 210 215 220 Ser Gly Ile Leu Lys Asp Phe Trp Gly Asn Tyr Leu Gln Tyr Asp Lys 225 230 235 240 Pro Tyr Tyr Met Leu Asn Leu Phe Asp Pro Asn Lys Tyr Val Asp Val 245 250 255 Asn Asn Ile Gly Ile Arg Gly Tyr Met Tyr Leu Lys Gly Pro Arg Gly 260 265 270 Ser Val Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr Leu Tyr Glu Gly 275 280 285 Thr Lys Phe Ile Ile Lys Lys Tyr Ala Ser Gly Asn Glu Asp Asn Ile 290 295 300 Val Arg Asn Asn Asp Arg Val Tyr Ile Asn Val Val Val Lys Asn Lys 305 310 315 320 Glu Tyr Arg Leu Ala Thr Asn Ala Ser Gln Ala Gly Val Glu Lys Ile 325 330 335 Leu Ser Ala Leu Glu Ile Pro Asp Val Gly Asn Leu Ser Gln Val Val 340 345 350 Val Met Lys Ser Lys Asp Asp Gln Gly Ile Arg Asn Lys Cys Lys Met 355 360 365 Asn Leu Gln Asp Asn Asn Gly Asn Asp Ile Gly Phe Ile Gly Phe His 370 375 380 Leu Tyr Asp Asn Ile Ala Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg 385 390 395 400 Gln Val Gly Lys Ala Ser Arg Thr Phe Gly Cys Ser Trp Glu Phe Ile 405 410 415 Pro Val Asp Asp Gly Trp Gly Glu Ser Ser Leu 420 425 <210> 101 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 101 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Arg Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 102 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype RBD <400> 102 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile 1 5 10 15 Gly Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Glu Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Leu Tyr Asp Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 103 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype RBD <400> 103 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Arg Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 104 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A5 subtype RBD <400> 104 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Glu Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Ser Lys Ile Glu Ile Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Ala Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Ile Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Ile Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Val Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Asp Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 105 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A3 subtype RBD <400> 105 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile 1 5 10 15 Gly Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Met Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Pro Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Val Gly Phe His Leu Tyr Asp Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 106 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 106 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile 1 5 10 15 Gly Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Arg Val Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Glu Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Leu Tyr Asp Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 107 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 107 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile 1 5 10 15 Gly Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Glu Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Leu Tyr Asp Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 108 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 108 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Lys Ile Asn Ile 1 5 10 15 Gly Asp Arg Val Tyr Tyr Asp Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Glu Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Leu Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 109 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype RBD <400> 109 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Ile Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Asn Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 110 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype RBD <400> 110 Met Asn Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn 1 5 10 15 Ile Gly Ser Arg Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln 20 25 30 Leu Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala 35 40 45 Ile Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile 50 55 60 Lys Ile Pro Lys Tyr Phe Ser Glu Ile Ser Leu Asn Asn Glu Tyr Thr 65 70 75 80 Ile Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn 85 90 95 Tyr Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln 100 105 110 Arg Val Val Phe Lys Tyr Ser Gln Met Val Ala Ile Ser Asp Tyr Ile 115 120 125 Asn Arg Trp Ile Phe Ile Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser 130 135 140 Lys Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn 145 150 155 160 Leu Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly 165 170 175 Cys Arg Asp Pro Arg Arg Tyr Ile Met Ile Lys Tyr Phe Asn Leu Phe 180 185 190 Asp Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp 195 200 205 Phe Trp Gly Asn Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn 210 215 220 Leu Phe Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg 225 230 235 240 Gly Tyr Met Tyr Leu Lys Gly Ser Arg Ser Thr Leu Leu Thr Thr Asn 245 250 255 Ile Tyr Leu Asn Ser Gly Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys 260 265 270 Lys Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg 275 280 285 Val Tyr Ile Asn Val Val Val Asn Asn Lys Glu Tyr Arg Leu Ala Thr 290 295 300 Asn Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile 305 310 315 320 Pro Asp Ile Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn 325 330 335 Asp Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn 340 345 350 Gly Asn Asp Ile Gly Phe Ile Gly Phe His Lys Phe Asn Asp Ile Tyr 355 360 365 Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Ile Ser Ser 370 375 380 Arg Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp 385 390 395 400 Gly Glu <210> 111 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 111 Met Asp Asp Leu Ile Asp Leu Ser Arg Tyr Gly Ala Glu Ile Tyr Asn 1 5 10 15 Gly Asp Lys Val Tyr Tyr Asn Ser Ile Asp Lys Asn Gln Ile Arg Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Lys Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Phe Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Ile Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro His Arg Tyr Ile Val Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Ser Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Asp Asn Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Arg 385 390 395 400 Glu <210> 112 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A1 subtype RBD <400> 112 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Arg Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 113 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 113 Met Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile 1 5 10 15 Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu 20 25 30 Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg 50 55 60 Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Ile Asp Val Asn Asn Val Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Asn Val Met Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Ser Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 325 330 335 Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 370 375 380 Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 114 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A-A2 subtype RBD <400> 114 Met Asp Glu Leu Ile Asp Leu Ser Arg Tyr Gly Ala Glu Ile Tyr Arg 1 5 10 15 Gly Asp Lys Val Phe Tyr Asn Ser Ile Asp Lys Asn Gln Ile Lys Leu 20 25 30 Ile Asn Leu Glu Ser Ser Thr Ile Glu Val Ile Leu Lys Asn Ala Ile 35 40 45 Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Lys 50 55 60 Ile Pro Lys Tyr Phe Ser Lys Ile Asn Leu Asn Asn Glu Tyr Thr Ile 65 70 75 80 Ile Asn Cys Ile Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr 85 90 95 Gly Glu Ile Ile Trp Thr Leu Gln Asp Asn Lys Gln Asn Ile Gln Arg 100 105 110 Val Val Phe Lys Tyr Ser Gln Met Val Asn Ile Ser Asp Tyr Ile Asn 115 120 125 Arg Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Thr Lys Ser Lys 130 135 140 Ile Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Leu Ile Ser Asn Leu 145 150 155 160 Gly Asn Ile His Ala Ser Asn Lys Ile Met Phe Lys Leu Asp Gly Cys 165 170 175 Arg Asp Pro Gly Arg Tyr Ile Val Ile Lys Tyr Phe Asn Leu Phe Asp 180 185 190 Lys Glu Leu Tyr Asp Ser Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 195 200 205 Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 210 215 220 Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Ile Gly Ile Arg Gly 225 230 235 240 Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Val Thr Thr Asn Ile 245 250 255 Tyr Leu Asn Ser Thr Leu Tyr Met Gly Thr Lys Phe Ile Ile Lys Lys 260 265 270 Tyr Ala Ser Gly Asn Glu Asp Asn Ile Val Arg Asn Asn Asp Arg Val 275 280 285 Tyr Ile Asn Val Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 290 295 300 Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 305 310 315 320 Asp Val Gly Asn Leu Ser Gln Leu Val Val Met Lys Ser Lys Asp Asp 325 330 335 Gln Gly Ile Arg Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 340 345 350 Asn Asp Ile Gly Leu Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 355 360 365 Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Val Gly Lys Ala Ser Arg 370 375 380 Thr Phe Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 385 390 395 400 Glu <210> 115 <211> 425 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A RBD <400> 115 Met Ile Ile Asn Thr Ser Ile Leu Asn Leu Arg Tyr Glu Ser Asn His 1 5 10 15 Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile Gly Ser Lys 20 25 30 Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu Phe Asn Leu 35 40 45 Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile Val Tyr Asn 50 55 60 Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg Ile Pro Lys 65 70 75 80 Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile Ile Asn Cys 85 90 95 Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr Gly Glu Ile 100 105 110 Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg Val Val Phe 115 120 125 Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn Arg Trp Ile 130 135 140 Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys Ile Tyr Ile 145 150 155 160 Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu Gly Asn Ile 165 170 175 His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys Arg Asp Thr 180 185 190 His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp Lys Glu Leu 195 200 205 Asn Glu Lys Glu Ile Lys Asp Leu Tyr Asp Asn Gln Ser Asn Ser Gly 210 215 220 Ile Leu Lys Asp Phe Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr 225 230 235 240 Tyr Met Leu Asn Leu Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn 245 250 255 Val Gly Ile Arg Gly Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val 260 265 270 Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser Leu Tyr Arg Gly Thr Lys 275 280 285 Phe Ile Ile Lys Lys Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg 290 295 300 Asn Asn Asp Arg Val Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr 305 310 315 320 Arg Leu Ala Thr Asn Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser 325 330 335 Ala Leu Glu Ile Pro Asp Val Gly Asn Leu Ser Gln Val Val Val Met 340 345 350 Lys Ser Lys Asn Asp Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu 355 360 365 Gln Asp Asn Asn Gly Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe 370 375 380 Asn Asn Ile Ala Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile 385 390 395 400 Glu Arg Ser Ser Arg Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val 405 410 415 Asp Asp Gly Trp Gly Glu Arg Pro Leu 420 425 <210> 116 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A1-A2 <400> 116 Arg Gly Ser Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser 1 5 10 <210> 117 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A2-A2 <400> 117 Arg Gly Ser Val Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr 1 5 10 <210> 118 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A3-A2 <400> 118 Arg Gly Ser Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Thr 1 5 10 <210> 119 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A4-A2 <400> 119 Arg Gly Asp Asn Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser 1 5 10 <210> 120 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> BoNT/A5-A2 <400> 120 Arg Gly Ser Val Ile Val Thr Asn Ile Tyr Leu Asn Ser Ser 1 5 10 <210> 121 <211> 437 <212> PRT <213> Artificial Sequence <220> <223> BoNTs LC/amino acid sequence 1 <400> 121 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe 435 <210> 122 <211> 451 <212> PRT <213> Artificial Sequence <220> <223> BoNTs LC-A2/amino acid sequence 2 <400> 122 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Arg Gly Ser Val Val Thr Thr Asn Ile Tyr Leu 435 440 445 Asn Ser Thr 450 <210> 123 <211> 464 <212> PRT <213> Artificial Sequence <220> <223> BoNTs LC-Aurein 1.2-A2/amino acid sequence 3 <400> 123 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe Gly Leu Phe Asp Ile Ile Lys Lys Ile Ala Glu 435 440 445 Ser Phe Arg Gly Ser Val Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr 450 455 460 <210> 124 <211> 461 <212> PRT <213> Artificial Sequence <220> <223> Recombinant botulinum neurotoxin amino acid sequence 4 <400> 124 Met Pro Phe Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly 1 5 10 15 Val Asp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro 20 25 30 Val Lys Ala Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg 35 40 45 Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu 50 55 60 Ala Lys Gln Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr 65 70 75 80 Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu 85 90 95 Arg Ile Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val 100 105 110 Arg Gly Ile Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys 115 120 125 Val Ile Asp Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr 130 135 140 Arg Ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile 145 150 155 160 Ile Gln Phe Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr 165 170 175 Arg Asn Gly Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe 180 185 190 Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu 195 200 205 Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu 210 215 220 Leu Ile His Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn 225 230 235 240 Arg Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250 255 Glu Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265 270 Phe Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280 285 Lys Phe Lys Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile 290 295 300 Gly Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys 305 310 315 320 Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu 325 330 335 Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp 340 345 350 Asn Phe Val Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn 355 360 365 Phe Asp Lys Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr 370 375 380 Thr Ile Lys Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn 385 390 395 400 Phe Asn Gly Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu 405 410 415 Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg 420 425 430 Gly Ile Ile Pro Phe His His His His His His His His His His Arg 435 440 445 Gly Ser Val Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr 450 455 460 <210> 125 <211> 474 <212> PRT <213> Artificial Sequence <220> <223> Aurein 1.2-BoNTs LC-His10-A2/amino acid sequence 5 <400> 125 Met Gly Leu Phe Asp Ile Ile Lys Lys Ile Ala Glu Ser Phe Pro Phe 1 5 10 15 Val Asn Lys Gln Phe Asn Tyr Lys Asp Pro Val Asn Gly Val Asp Ile 20 25 30 Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gln Met Gln Pro Val Lys Ala 35 40 45 Phe Lys Ile His Asn Lys Ile Trp Val Ile Pro Glu Arg Asp Thr Phe 50 55 60 Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Glu Ala Lys Gln 65 70 75 80 Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr Asp Asn Glu 85 90 95 Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu Arg Ile Tyr 100 105 110 Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser Ile Val Arg Gly Ile 115 120 125 Pro Phe Trp Gly Gly Ser Thr Ile Asp Thr Glu Leu Lys Val Ile Asp 130 135 140 Thr Asn Cys Ile Asn Val Ile Gln Pro Asp Gly Ser Tyr Arg Ser Glu 145 150 155 160 Glu Leu Asn Leu Val Ile Ile Gly Pro Ser Ala Asp Ile Ile Gln Phe 165 170 175 Glu Cys Lys Ser Phe Gly His Asp Val Leu Asn Leu Thr Arg Asn Gly 180 185 190 Tyr Gly Ser Thr Gln Tyr Ile Arg Phe Ser Pro Asp Phe Thr Phe Gly 195 200 205 Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu Gly Ala Gly 210 215 220 Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala His Glu Leu Ile His 225 230 235 240 Ala Glu His Arg Leu Tyr Gly Ile Ala Ile Asn Pro Asn Arg Val Phe 245 250 255 Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu Glu Val Ser 260 265 270 Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys Phe Ile Asp 275 280 285 Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn Lys Phe Lys 290 295 300 Asp Val Ala Ser Thr Leu Asn Lys Ala Lys Ser Ile Ile Gly Thr Thr 305 310 315 320 Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys Tyr Leu Leu 325 330 335 Ser Glu Asp Thr Ser Gly Lys Phe Ser Val Asp Lys Leu Lys Phe Asp 340 345 350 Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp Asn Phe Val 355 360 365 Asn Phe Phe Lys Val Ile Asn Arg Lys Thr Tyr Leu Asn Phe Asp Lys 370 375 380 Ala Val Phe Arg Ile Asn Ile Val Pro Asp Glu Asn Tyr Thr Ile Lys 385 390 395 400 Asp Gly Phe Asn Leu Lys Gly Ala Asn Leu Ser Thr Asn Phe Asn Gly 405 410 415 Gln Asn Thr Glu Ile Asn Ser Arg Asn Phe Thr Arg Leu Lys Asn Phe 420 425 430 Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Arg Gly Ile Ile 435 440 445 Pro Phe His His His His His His His His His His His Arg Gly Ser Val 450 455 460 Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr 465 470 <210> 126 <211> 474 <212> PRT <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/amino acid sequence 6 <400> 126 Met Gly Leu Phe Asp Ile Ile Lys Lys Ile Ala Glu Ser Phe His His 1 5 10 15 His His His His His His His His Pro Phe Val Asn Lys Gln Phe Asn 20 25 30 Tyr Lys Asp Pro Val Asn Gly Val Asp Ile Ala Tyr Ile Lys Ile Pro 35 40 45 Asn Ala Gly Gln Met Gln Pro Val Lys Ala Phe Lys Ile His Asn Lys 50 55 60 Ile Trp Val Ile Pro Glu Arg Asp Thr Phe Thr Asn Pro Glu Glu Gly 65 70 75 80 Asp Leu Asn Pro Pro Pro Glu Ala Lys Gln Val Pro Val Ser Tyr Tyr 85 90 95 Asp Ser Thr Tyr Leu Ser Thr Asp Asn Glu Lys Asp Asn Tyr Leu Lys 100 105 110 Gly Val Thr Lys Leu Phe Glu Arg Ile Tyr Ser Thr Asp Leu Gly Arg 115 120 125 Met Leu Leu Thr Ser Ile Val Arg Gly Ile Pro Phe Trp Gly Gly Ser 130 135 140 Thr Ile Asp Thr Glu Leu Lys Val Ile Asp Thr Asn Cys Ile Asn Val 145 150 155 160 Ile Gln Pro Asp Gly Ser Tyr Arg Ser Glu Glu Leu Asn Leu Val Ile 165 170 175 Ile Gly Pro Ser Ala Asp Ile Ile Gln Phe Glu Cys Lys Ser Phe Gly 180 185 190 His Asp Val Leu Asn Leu Thr Arg Asn Gly Tyr Gly Ser Thr Gln Tyr 195 200 205 Ile Arg Phe Ser Pro Asp Phe Thr Phe Gly Phe Glu Glu Ser Leu Glu 210 215 220 Val Asp Thr Asn Pro Leu Leu Gly Ala Gly Lys Phe Ala Thr Asp Pro 225 230 235 240 Ala Val Thr Leu Ala His Glu Leu Ile His Ala Glu His Arg Leu Tyr 245 250 255 Gly Ile Ala Ile Asn Pro Asn Arg Val Phe Lys Val Asn Thr Asn Ala 260 265 270 Tyr Tyr Glu Met Ser Gly Leu Glu Val Ser Phe Glu Glu Leu Arg Thr 275 280 285 Phe Gly Gly His Asp Ala Lys Phe Ile Asp Ser Leu Gln Glu Asn Glu 290 295 300 Phe Arg Leu Tyr Tyr Tyr Asn Lys Phe Lys Asp Val Ala Ser Thr Leu 305 310 315 320 Asn Lys Ala Lys Ser Ile Ile Gly Thr Thr Ala Ser Leu Gln Tyr Met 325 330 335 Lys Asn Val Phe Lys Glu Lys Tyr Leu Leu Ser Glu Asp Thr Ser Gly 340 345 350 Lys Phe Ser Val Asp Lys Leu Lys Phe Asp Lys Leu Tyr Lys Met Leu 355 360 365 Thr Glu Ile Tyr Thr Glu Asp Asn Phe Val Asn Phe Phe Lys Val Ile 370 375 380 Asn Arg Lys Thr Tyr Leu Asn Phe Asp Lys Ala Val Phe Arg Ile Asn 385 390 395 400 Ile Val Pro Asp Glu Asn Tyr Thr Ile Lys Asp Gly Phe Asn Leu Lys 405 410 415 Gly Ala Asn Leu Ser Thr Asn Phe Asn Gly Gln Asn Thr Glu Ile Asn 420 425 430 Ser Arg Asn Phe Thr Arg Leu Lys Asn Phe Thr Gly Leu Phe Glu Phe 435 440 445 Tyr Lys Leu Leu Cys Val Arg Gly Ile Ile Pro Phe Arg Gly Ser Val 450 455 460 Val Thr Thr Asn Ile Tyr Leu Asn Ser Thr 465 470 <210> 127 <211> 1311 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC/DNA sequence 1 <400> 127 atgcccttcg tcaacaaaca gttcaattat aaggatccgg taaatggggt agacatagca 60 tacataaaga ttcccaacgc aggacaaatg caaccggtga aggcgttcaa aattcataat 120 aagatatggg tcatacctga gcgggacacc tttacaaacc ccgaggaagg tgatcttaac 180 ccgcctccgg aagcgaaaca agttcctgtt tcctactatg actcaactta cctgtcaact 240 gataatgaga aggacaatta tttaaaaggc gtcacgaagt tattcgaaag aatatattca 300 actgacctgg gacgcatgct gctgacttct atcgttagag ggataccatt ctggggaggt 360 tctactattg atactgagct gaaggtaatt gacaccaact gtattaatgt tattcaacct 420 gatggttcct acagatcaga agagttaaat cttgtcatta taggtcccag tgcagacatc 480 attcagtttg aatgtaaatc tttcgggcac gatgtgctta atttgaccag aaatggttat 540 ggaagtactc aatacattag attttcgccg gactttacat tcggctttga ggaaagtctt 600 gaagtggaca cgaacccttt gctgggggca ggtaaatttg ctacggatcc agcggtcacc 660 cttgcacacg agcttatcca tgcggagcac cgcttatacg gcatagcaat aaaccccaat 720 cgtgtcttta aggtaaatac gaacgcttac tatgaaatgt ctggtctgga ggtaagcttt 780 gaagaactgc gtactttcgg ggggcacgac gccaagttca tcgactccct gcaagagaat 840 gagtttcggc tgtactatta caataaattc aaggacgtag cgagcacctt aaataaagcc 900 aaaagcatca ttggcactac tgcctcttta cagtacatga agaacgtatt taaagagaag 960 tatcttttaa gcgaagatac gtcgggcaag tttagcgtcg ataagttaaa gtttgacaag 1020 ttatataaga tgttgacgga gatttataca gaggataact tcgttaactt cttcaaagta 1080 ataaaccgta aaacgtacct gaacttcgat aaggctgtgt tccgtataaa catcgtgccg 1140 gacgagaact acacaattaa ggacggcttt aacttaaaag gggcaaacct gagtaccaac 1200 ttcaacggcc aaaatactga gataaattca cgtaatttta cccgtttgaa gaattttacg 1260 ggcctgttcg agttttacaa gttactttgt gtccgtggca tcattccctt c 1311 <210> 128 <211> 1353 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-A2/DNA sequence 2 <400> 128 atgcctttcg ttaataagca gttcaactat aaggatcctg tcaacggtgt cgacatcgcg 60 tacataaaaa tcccgaatgc tggtcaaatg cagccggtaa aagccttcaa aattcacaat 120 aagatctggg taatacctga gcgtgatacg ttcacgaatc cagaggaagg tgatttgaac 180 ccgcctccag aggcaaaaca ggttcctgta tcgtattacg atagtaccta cctgagtacg 240 gacaacgaga aagacaatta tttaaagggt gtcacgaaac tgtttgagcg gatttactca 300 acggatttgg gaagaatgct gttgacgtca atcgtaagag gtattccttt ctggggtggt 360 tcaaccatag atacagaact gaaggtcatt gatactaatt gtataaatgt catacagcct 420 gatggttctt atcgctccga agaactgaac ttggttatta tagggccatc cgctgatata 480 attcagtttg agtgtaaaag tttcggccac gacgtcttaa acttgacaag aaacgggtac 540 gggtctactc aatatattcg cttcagccct gatttcactt ttggcttcga ggaatcgtta 600 gaagtggaca caaatccgct tctgggggcg ggaaaatttg ctacggaccc cgcggtcacg 660 cttgctcacg agttaattca tgcagagcat cggttatacg gcatcgccat caatccaaac 720 cgggttttca aggtaaatac gaacgcctac tacgaaatga gcggattaga ggtatcgttc 780 gaagaacttc ggacgtttgg cggacatgac gccaaattca tagattcact gcaagaaaat 840 gaatttagat tgtattatta caacaagttt aaagacgtgg cgtccaccct taataaggcc 900 aagtctatca taggtacaac agcatcgtta cagtatatga agaacgtttt caaggaaaaa 960 taccttttgt ctgaagacac cagcgggaag ttctcggttg ataaactgaa gtttgacaaa 1020 ctgtacaaga tgttgacaga aatctacact gaggacaatt ttgtcaattt ttttaaggtc 1080 attaatcgta agacatatct gaatttcgac aaagcggtat tccgtataaa catcgtgcca 1140 gatgagaatt atacaattaa ggacggcttc aacttaaaag gtgctaacct tagcaccaac 1200 tttaacggcc agaataccga gattaattcc cgcaacttca cccgcttaaa aaactttaca 1260 ggactttttg aattttacaa attgctgtgt gtgcggggca taatcccttt tagagggtct 1320 gtggtcacca caaatatata cctgaacagt act 1353 <210> 129 <211> 1392 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-Aurein 1.2-A2/DNA sequence 3 <400> 129 atgcctttcg taaacaaaca attcaactat aaagaccctg tcaatggggt agacatcgcg 60 tacataaaga ttcccaatgc gggccagatg caaccagtca aagcctttaa gatccacaac 120 aagatctggg tgattccaga acgcgacacg ttcaccaatc ctgaagaagg cgacttgaat 180 ccaccgccag aggccaaaca agtaccggta agctattacg actccaccta tttatcgaca 240 gataatgaga aagacaacta cttaaaaggt gtaaccaagt tatttgagcg tatatactcg 300 actgatctgg gaagaatgct gttaacttct attgttagag gaattccttt ttggggtgga 360 tccactatcg acaccgagtt aaaagttatt gacacaaatt gtataaacgt tatccagccc 420 gacggttcct accgttccga agaactgaac cttgtcatca ttggtccctc ggcagatata 480 attcaatttg aatgtaagag tttcgggcac gatgtcttga atctgactag aaacgggtat 540 ggatcaacac agtatattag attttcgccc gacttcacgt tcggattcga ggaaagttta 600 gaagtcgata ctaacccctt gttaggtgcg gggaaatttg caaccgaccc tgctgttacc 660 ttggcccacg agttgatcca cgcagagcat cggctttatg gaattgctat taacccaaac 720 cgcgtcttta aagtaaacac taacgcctac tatgagatga gtggcttgga ggtgagtttt 780 gaagagctta gaacctttgg tggccatgat gctaagttta tcgactcttt acaggaaaat 840 gagttcagat tgtactacta taacaaattt aaagacgttg catcaacttt gaataaagcc 900 aaatctataa ttgggactac tgcgtccctg caatacatga aaaacgtgtt caaggagaag 960 taccttctgt cggaagatac atctggtaaa tttagcgttg acaagttaaa gttcgataaa 1020 ctgtacaaga tgttaacaga aatatatacc gaggataatt tcgtcaactt tttcaaagtc 1080 attaatcgga agacttactt aaacttcgat aaagccgtct tccgcattaa cattgtgcca 1140 gacgagaact atacaattaa ggatggtttc aatttgaaag gagctaactt aagcactaac 1200 tttaacggcc agaacacaga gatcaattca cgtaacttca caagacttaa gaattttaca 1260 gggctgttcg agttttataa gctgctttgc gtaagaggaa tcatcccctt tggattattt 1320 gacattatta agaagatcgc cgaaagtttt cgtgggtccg ttgttacgac aaacatttat 1380 ttaaattcga ca 1392 <210> 130 <211> 1383 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-BoNTs LC-His10-A2/DNA sequence 4 <400> 130 atgccgtttg tgaacaaaca attcaactat aaagatccgg taaacggagt tgacatcgcg 60 tacatcaaaa taccgaatgc tggtcagatg cagccagtaa aagctttcaa aatccacaac 120 aaaatctggg tcatccctga aagagatacc tttacaaatc ccgaggaggg ggacctgaat 180 ccaccaccag aggcgaaaca ggtcccggtt tcatactatg acagtaccta cctttctact 240 gataacgaga aagataacta tttgaagggt gtaactaaat tatttgaaag aatttacagc 300 acggatttgg gtcggatgct gttaacatca attgtgcgcg gcataccatt ctggggagga 360 tcgactatag acaccgagtt aaaggtgatt gatactaact gtataaatgt aatacaaccc 420 gacggttcat acagaagtga agagttaaat cttgtcatca ttggtccttc agctgatatt 480 atacagtttg aatgcaaaag ttttggacac gacgtactta acttgacgcg taatggttac 540 ggcagtacac agtacatacg cttcagccct gacttcacgt tcggtttcga ggaaagtctt 600 gaagttgata ctaaccccct gcttggcgca ggcaaattcg ccactgatcc cgctgtcaca 660 ctggctcacg aactgataca tgcagagcat agactttacg gcatcgcaat taacccaaac 720 cgggtcttta aggtaaatac gaatgcttac tacgaaatgt ctggcctgga agtatcattc 780 gaggagttga gaaccttcgg cggccacgac gctaaattta tcgacagtct tcaggaaaac 840 gagttccgtc tgtactacta caataaattt aaggatgtgg cttctacttt gaataaggcc 900 aaaagtatca tcgggactac tgcatccttg caatatatga agaacgtatt caaggaaaaa 960 tacttgttat cagaggacac gagtggcaag ttctctgttg acaagttaaa attcgataaa 1020 ttatacaaga tgttgactga gatatacaca gaggataact ttgtcaattt ttttaaggtt 1080 ataaatcgga agacctattt gaacttcgat aaggctgtct ttcgtattaa tatagtgccc 1140 gatgaaaact ataccatcaa agacggtttc aatctgaaag gtgccaattt gtcgaccaac 1200 ttcaacgggc aaaacacgga gatcaatagt agaaatttca cgcgtcttaa aaacttcact 1260 ggtctgttcg aattttataa gcttttgtgc gttcgcggaa taattccttt tcatcatcat 1320 catcaccatc atcaccacca tcgtgggtcc gtagtgacca ctaacatcta cttaaattct 1380 acc 1383 <210> 131 <211> 1422 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-BoNTs LC-His10-A2/DNA sequence 5 <400> 131 atggggttat tcgacataat caaaaagata gctgaatcct ttccgttcgt aaacaagcag 60 ttcaattata aagatccagt gaacggggtg gacattgcct acattaagat acccaatgcg 120 ggccagatgc aacccgtaaa agcgttcaaa atacacaaca aaatatgggt gattcctgag 180 cgcgacacat ttacgaaccc ggaagaaggt gatttgaatc cgccacctga agcgaaacag 240 gtccccgtga gttattacga tagcacgtac ttatcaactg ataatgagaa agataactat 300 ttgaagggcg tgacgaagtt attcgagcgc atatactcta ccgatcttgg tcggatgctg 360 ttaacgagca tcgtacgggg cattcccttc tggggtgggt ccacaatcga tacggagtta 420 aaagtcatcg acaccaactg tatcaacgtc atccagcctg acgggtcgta cagaagtgaa 480 gagttaaact tggttataat aggaccatcc gcagatatta tacaattcga gtgcaagtct 540 tttggtcatg atgttttgaa cctgacgcgc aacggatatg gctccacgca atacatccgc 600 ttctccccag attttacatt tggattcgaa gaatcattag aagtagacac aaacccactg 660 ctgggggctg gcaaattcgc aactgacccc gcagttacct tagcgcacga actgatacac 720 gcggaacatc ggctgtatgg catagcgatt aaccccaacc gtgttttcaa ggtaaacacg 780 aatgcgtact acgagatgtc tggtttggaa gtttcttttg aggagctgcg cacctttggg 840 ggtcatgatg cgaaatttat tgacagcttg caagagaatg agttcagatt atattactac 900 aataaattta aggatgtggc gagcactctg aataaggcaa aatccattat cggtactact 960 gcatctttgc agtatatgaa aaatgtgttc aaggagaagt acttgttgtc tgaggatacg 1020 agtggtaaat tctcagttga caaattgaaa tttgataagc tgtataagat gttgacagag 1080 atttatacgg aggataactt tgtgaatttc ttcaaagtca taaaccgtaa aacctacctg 1140 aattttgata aggcggtgtt tcggattaat atagttccgg acgagaatta tacaatcaag 1200 gacggcttca atttaaaagg agctaattta tccaccaatt ttaacggcca aaacacagaa 1260 atcaactctc gtaatttcac gcgtttaaag aacttcactg ggctgttcga attttataaa 1320 ttgttatgtg tccgcggcat aataccgttt catcatcacc atcatcatca tcaccatcac 1380 cgtggatcag tggtcaccac taatatttac ctgaacagta cg 1422 <210> 132 <211> 1422 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/DNA sequence 6 <400> 132 atgggtcttt tcgacataat aaagaaaatc gccgaaagct ttcatcacca tcaccatcat 60 catcatcacc atccgttcgt aaataagcag tttaattaca aagatcccgt gaacggcgtg 120 gacatcgcct atataaaaat cccgaatgcc ggccagatgc agcccgttaa ggccttcaag 180 atacacaata aaatctgggt cataccggag cgggatacat tcacaaaccc ggaagaaggt 240 gaccttaacc ctcccccgga ggcaaaacaa gttcccgttt catattatga ctctacgtac 300 ctttccaccg acaacgaaaa ggacaattat ctgaaaggcg tgacaaaact tttcgaacgc 360 atttattcta cagatcttgg tagaatgctg ttgacttcta ttgtcagagg aattcctttc 420 tggggcggga gcactataga cactgagtta aaggtaattg acacgaattg cataaatgtt 480 atacaaccgg acggcagtta ccgttcggag gagcttaatt tagttataat cggtccatca 540 gctgacataa ttcaatttga gtgcaaatca tttggacacg atgttttaaa tttgactcgt 600 aatggatacg gctccacgca atacatccgt ttcagtcctg attttacatt cggttttgaa 660 gagtctcttg aggtggacac gaaccctttg ttaggcgccg gcaaatttgc tactgatcca 720 gccgtcacat tagctcacga acttattcac gccgaacacc gtctgtacgg tattgcaata 780 aatccaaacc gggtctttaa agtaaacacg aacgcctact acgaaatgtc gggtcttgag 840 gtctcattcg aggaattaag aacctttgga ggtcacgatg cgaagttcat tgactccttg 900 caagagaacg aatttagact ttactattac aacaagttca aggatgtcgc gtcgacctta 960 aataaggcca aatccataat tgggacgaca gcaagccttc aatacatgaa aaacgtcttc 1020 aaggagaagt atcttttaag cgaggacacc agcggtaagt ttagtgtcga taagttaaaa 1080 ttcgacaaat tgtacaagat gcttactgag atttacacag aagacaattt tgttaatttt 1140 ttcaaagtta taaatcgtaa aacctatctg aatttcgata aagcggtttt ccgtatcaac 1200 attgtgccgg acgagaatta caccataaaa gacgggttca atttaaaggg cgcaaacctt 1260 tctaccaatt ttaacggaca aaatacggaa atcaactcac gcaattttac gcgcctgaaa 1320 aattttacag gcctttttga attttataag ttgctttgtg tacggggtat tatacccttt 1380 cggggatcag tggtgactac taatatatac ttgaactcga ct 1422 <210> 133 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> pET28b/Forward primer <400> 133 caccaccacc accaccactg agatc 25 <210> 134 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> pET28b/Backward primer <400> 134 ctcgctaccg cctccaccac tg 22 <210> 135 <211> 48 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC/Forward primer <400> 135 gtggtggtgg tggtggaagg gaatgatgcc acggacacaa agtaactt 48 <210> 136 <211> 53 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC/Backward primer <400> 136 ggaggcggta gcgagatgcc cttcgtcaac aaacagttca attataagga tcc 53 <210> 137 <211> 76 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-A2/Forward primer <400> 137 tgtgctgttc aaatagatgt ttgtggtcac aacgctcccg cgagagggtc tgtggtcacc 60 acaaatatat acctga 76 <210> 138 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-A2/Backward primer <400> 138 caccaccacc accaccactg agatc 25 <210> 139 <211> 72 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-Aurein 1.2-A2/Forward primer <400> 139 aaatgattct gcgatcttta ttatgatgtc aaacagtcca gagggtctgt ggtcaccaca 60 aatatatacc tg 72 <210> 140 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-Aurein 1.2-A2/Backward primer <400> 140 cgcgggagcg ttgtgaccac aaacat 26 <210> 141 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-His10-A2/ Forward primer <400> 141 gtgatggtga tggtgatggt gatggtggtg agagggtctg tggtcaccac aaatatatac 60 ctg 63 <210> 142 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> BoNTs LC-His10-A2/Backward primer <400> 142 cgcgggagcg ttgtgaccac aaacat 26 <210> 143 <211> 58 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-BoNTs LC-His10-A2/Forward primer <400> 143 aaatgattct gcgatcttta ttatgatgtc aaacagctcg ctaccgcctc caccactg 58 <210> 144 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-BoNTs LC-His10-A2/Backward primer <400> 144 atgcccttcg tcaacaaaca gttcaattat aaggatcc 38 <210> 145 <211> 52 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/Backward primer <400> 145 gtgatggtga tggtgatggt gatggtggtg ctcgctaccg cctccaccac tg 52 <210> 146 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/Backward primer <400> 146 atgcccttcg tcaacaaaca gttcaattat aaggatcc 38 <210> 147 <211> 61 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/Forward primer <400> 147 aaatgattct gcgatcttta ttatgatgtc aaacagtccc tcgctaccgc ctccaccact 60 g 61 <210> 148 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> Aurein 1.2-His10-BoNTs LC-A2/Backward primer <400> 148 caccaccatc accatcacca tcaccatc 28 <210> 149 <211> 427 <212> PRT <213> Artificial Sequence <220> <223> RBD <400> 149 Met Lys Asn Ile Ile Asn Thr Ser Ile Leu Asn Leu Arg Tyr Glu Ser 1 5 10 15 Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile Gly 20 25 30 Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu Phe 35 40 45 Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile Val 50 55 60 Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg Ile 65 70 75 80 Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile Ile 85 90 95 Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr Gly 100 105 110 Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg Val 115 120 125 Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn Arg 130 135 140 Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys Ile 145 150 155 160 Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu Gly 165 170 175 Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys Arg 180 185 190 Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp Lys 195 200 205 Glu Leu Asn Glu Lys Glu Ile Lys Asp Leu Tyr Asp Asn Gln Ser Asn 210 215 220 Ser Gly Ile Leu Lys Asp Phe Trp Gly Asp Tyr Leu Gln Tyr Asp Lys 225 230 235 240 Pro Tyr Tyr Met Leu Asn Leu Tyr Asp Pro Asn Lys Tyr Ile Asp Val 245 250 255 Asn Asn Val Gly Ile Arg Gly Tyr Met Tyr Leu Lys Gly Pro Arg Gly 260 265 270 Asn Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser Leu Tyr Met Gly 275 280 285 Thr Lys Phe Ile Ile Lys Lys Tyr Ala Ser Gly Asn Lys Asp Asn Ile 290 295 300 Val Arg Asn Asn Asp Arg Val Tyr Ile Asn Val Val Val Lys Asn Lys 305 310 315 320 Glu Tyr Arg Leu Ala Thr Asn Ala Ser Gln Ala Gly Val Glu Lys Ile 325 330 335 Leu Ser Ala Leu Glu Ile Pro Asp Val Gly Asn Leu Ser Gln Val Val 340 345 350 Val Met Lys Ser Lys Asn Asp Gln Gly Ile Thr Asn Lys Cys Lys Met 355 360 365 Asn Leu Gln Asp Asn Asn Gly Asn Asp Ile Gly Phe Ile Gly Phe His 370 375 380 Gln Phe Asn Asn Ile Ala Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg 385 390 395 400 Gln Ile Glu Arg Ser Ser Arg Thr Leu Gly Cys Ser Trp Glu Phe Ile 405 410 415 Pro Val Asp Asp Gly Trp Gly Glu Arg Pro Leu 420 425 <210> 150 <211> 427 <212> PRT <213> Artificial Sequence <220> <223> RBD' <400> 150 Met Lys Asn Ile Ile Asn Thr Ser Ile Leu Asn Leu Arg Tyr Glu Ser 1 5 10 15 Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser Lys Ile Asn Ile Gly 20 25 30 Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn Gln Ile Gln Leu Phe 35 40 45 Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu Lys Asn Ala Ile Val 50 55 60 Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser Phe Trp Ile Arg Ile 65 70 75 80 Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn Glu Tyr Thr Ile Ile 85 90 95 Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val Ser Leu Asn Tyr Gly 100 105 110 Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu Ile Lys Gln Arg Val 115 120 125 Val Phe Lys Tyr Ser Gln Met Ile Asn Ile Ser Asp Tyr Ile Asn Arg 130 135 140 Trp Ile Phe Val Thr Ile Thr Asn Asn Arg Leu Asn Asn Ser Lys Ile 145 150 155 160 Tyr Ile Asn Gly Arg Leu Ile Asp Gln Lys Pro Ile Ser Asn Leu Gly 165 170 175 Asn Ile His Ala Ser Asn Asn Ile Met Phe Lys Leu Asp Gly Cys Arg 180 185 190 Asp Thr His Arg Tyr Ile Trp Ile Lys Tyr Phe Asn Leu Phe Asp Lys 195 200 205 Glu Leu Asn Glu Lys Glu Ile Lys Asp Leu Tyr Asp Asn Gln Ser Asn 210 215 220 Ser Gly Ile Leu Lys Asp Phe Trp Gly Asp Tyr Leu Gln Tyr Asp Lys 225 230 235 240 Pro Tyr Tyr Met Leu Asn Leu Tyr Asp Pro Asn Lys Tyr Val Asp Val 245 250 255 Asn Asn Val Gly Ile Arg Gly Tyr Met Tyr Leu Lys Gly Pro Arg Gly 260 265 270 Ser Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser Leu Tyr Arg Gly 275 280 285 Thr Lys Phe Ile Ile Lys Lys Tyr Ala Ser Gly Asn Lys Asp Asn Ile 290 295 300 Val Arg Asn Asn Asp Arg Val Tyr Ile Asn Val Val Val Lys Asn Lys 305 310 315 320 Glu Tyr Arg Leu Ala Thr Asn Ala Ser Gln Ala Gly Val Glu Lys Ile 325 330 335 Leu Ser Ala Leu Glu Ile Pro Asp Val Gly Asn Leu Ser Gln Val Val 340 345 350 Val Met Lys Ser Lys Asn Asp Gln Gly Ile Thr Asn Lys Cys Lys Met 355 360 365 Asn Leu Gln Asp Asn Asn Gly Asn Asp Ile Gly Phe Ile Gly Phe His 370 375 380 Gln Phe Asn Asn Ile Ala Lys Leu Val Ala Ser Asn Trp Tyr Asn Arg 385 390 395 400 Gln Ile Glu Arg Ser Ser Arg Thr Leu Gly Cys Ser Trp Glu Phe Ile 405 410 415 Pro Val Asp Asp Gly Trp Gly Glu Arg Pro Leu 420 425 <210> 151 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> A2 <400> 151 Arg Gly Asn Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser 1 5 10 <210> 152 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> A2' <400> 152 Arg Gly Ser Val Met Thr Thr Asn Ile Tyr Leu Asn Ser Ser 1 5 10 <210> 153 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> SV2C <400> 153 Asp Val Ile Lys Pro Leu Gln Ser Asp Glu Tyr Ala Leu Leu Thr Arg 1 5 10 15 Asn Val Glu Arg Asp Lys Tyr Ala Asn Phe Thr Ile Asn Phe Thr Met 20 25 30 Glu Asn Gln Ile His Thr Gly Met Glu Tyr Asp Asn Gly Arg Phe Ile 35 40 45 Gly Val Lys Phe Lys Ser Val Thr Phe Lys Asp Ser Val Phe Lys Ser 50 55 60 Cys Thr Phe Glu Asp Val Thr Ser Val Asn Thr Tyr Phe Lys Asn Cys 65 70 75 80 Thr Phe Ile Asp Thr Val Phe Asp Asn Thr Asp Phe Glu Pro Tyr Lys 85 90 95 Phe Ile Asp Ser Glu Phe Lys Asn Cys Ser Phe Phe His Asn Lys Thr 100 105 110 Gly Cys Gln Ile Thr Phe Asp 115 <210> 154 <211> 1281 <212> DNA <213> Artificial Sequence <220> <223> RBD <400> 154 atgaaaaaca ttattaatac ttccatactt aacctgcgct acgagtctaa ccatttgatt 60 gacttatcgc ggtacgcttc taaaataaac atcgggagca aggtaaactt tgatccgata 120 gataaaaacc agattcaatt gttcaactta gagtctagta aaatagaggt tattttaaaa 180 aatgccattg tttataactc gatgtacgaa aatttctcga cctcgttttg gatacggata 240 ccgaagtact ttaactctat ttccttgaat aatgagtaca ccatcataaa ttgcatggaa 300 aacaattccg gttggaaagt gtcattaaac tatggcgaga taatttggac tttgcaggac 360 actcaggaga taaagcagcg tgtagtcttt aagtattccc agatgataaa catctcggac 420 tacataaatc gttggatttt tgtaaccata acaaataatc gccttaacaa tagcaaaatt 480 tatatcaacg gccggttgat cgatcaaaaa ccaatttcca acttggggaa catacacgct 540 tcgaataaca tcatgtttaa gcttgacggt tgtagagata cgcatcgcta catttggata 600 aagtatttta acctgtttga caaggagttg aatgagaaag aaattaagga cttatatgat 660 aatcaatcaa acagtgggat attgaaagat ttctggggcg actatcttca gtacgataag 720 ccctattata tgttgaactt atacgaccca aacaaataca ttgacgtaaa caatgtgggt 780 atcagaggat atatgtactt aaagggccca agagggaatg taatgacgac gaacatatac 840 cttaacagca gtctttacat gggcacgaaa ttcataataa aaaaatatgc gtccgggaat 900 aaggacaata ttgtaagaaa taacgaccgt gtatatatca atgtcgtggt caaaaacaaa 960 gagtaccggc tggctaccaa tgcttcccag gcaggtgtcg agaaaatact ttcggcgctt 1020 gaaattccag acgtgggcaa tctgtcacaa gtggtcgtta tgaaaagtaa aaatgatcaa 1080 gggatcacca ataagtgcaa gatgaatttg caagacaata atggtaacga tataggattt 1140 atagggtttc accagttcaa taacatcgct aagctggtcg ctagtaactg gtataataga 1200 caaattgaac ggtccagtcg cacgcttggc tgttcttggg agttcattcc ggttgatgat 1260 ggatggggcg aacgcccact g 1281 <210> 155 <211> 1281 <212> DNA <213> Artificial Sequence <220> <223> RBD' <400> 155 atgaaaaaca ttattaatac ttccatactt aacctgcgct acgagtctaa ccatttgatt 60 gacttatcgc ggtacgcttc taaaataaac atcgggagca aggtaaactt tgatccgata 120 gataaaaacc agattcaatt gttcaactta gagtctagta aaatagaggt tattttaaaa 180 aatgccattg tttataactc gatgtacgaa aatttctcga cctcgttttg gatacggata 240 ccgaagtact ttaactctat ttccttgaat aatgagtaca ccatcataaa ttgcatggaa 300 aacaattccg gttggaaagt gtcattaaac tatggcgaga taatttggac tttgcaggac 360 actcaggaga taaagcagcg tgtagtcttt aagtattccc agatgataaa catctcggac 420 tacataaatc gttggatttt tgtaaccata acaaataatc gccttaacaa tagcaaaatt 480 tatatcaacg gccggttgat cgatcaaaaa ccaatttcca acttggggaa catacacgct 540 tcgaataaca tcatgtttaa gcttgacggt tgtagagata cgcatcgcta catttggata 600 aagtatttta acctgtttga caaggagttg aatgagaaag aaattaagga cttatatgat 660 aatcaatcaa acagtgggat attgaaagat ttctggggcg actatcttca gtacgataag 720 ccctattata tgttgaactt atacgaccca aacaaatacg tggacgtaaa caatgtgggt 780 atcagaggat atatgtactt aaagggccca agagggtctg taatgacgac gaacatatac 840 cttaacagca gtctttaccg tggcacgaaa ttcataataa aaaaatatgc gtccgggaat 900 aaggacaata ttgtaagaaa taacgaccgt gtatatatca atgtcgtggt caaaaacaaa 960 gagtaccggc tggctaccaa tgcttcccag gcaggtgtcg agaaaatact ttcggcgctt 1020 gaaattccag acgtgggcaa tctgtcacaa gtggtcgtta tgaaaagtaa aaatgatcaa 1080 gggatcacca ataagtgcaa gatgaatttg caagacaata atggtaacga tataggattt 1140 atagggtttc accagttcaa taacatcgct aagctggtcg ctagtaactg gtataataga 1200 caaattgaac ggtccagtcg cacgcttggc tgttcttggg agttcattcc ggttgatgat 1260 ggatggggcg aacgcccact g 1281 <210> 156 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> A2 <400> 156 agagggaatg taatgacgac gaacatatac cttaacagca gt 42 <210> 157 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> A2' <400> 157 gggtctgtaa tgacgacgaa catatacctt aacagcagtc tt 42 <210> 158 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> SV2C <400> 158 gacgtgataa agccccttca gtcagatgaa tatgcattgc tgactcgcaa cgtggaacgt 60 gacaagtatg ccaattttac cataaatttc acaatggaaa accagatcca tacgggcatg 120 gagtacgaca acggtcggtt tatcggggta aaattcaagt cagtaacctt taaggattcg 180 gtgttcaaaa gttgcacatt tgaggacgtt acctccgtaa acacctactt caaaaattgc 240 acttttattg atacagtatt cgacaacacc gatttcgagc catataaatt catcgacagt 300 gagttcaaga actgcagttt ctttcataat aagaccggct gccagattac tttcgat 357 <210> 159 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> pET28b-EGFP/Forward primer <400> 159 gctaccgcct ccaccgtggt g 21 <210> 160 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> pET28b-EGFP/Backward primer <400> 160 tgagatccgg ctgctaacaa agcccgaaa 29 <210> 161 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> RBD insert/forward primer <400> 161 agcagccgga tctcacagtg ggcgttcgcc ccatccat 38 <210> 162 <211> 62 <212> DNA <213> Artificial Sequence <220> <223> RBD insert/Backward primer <400> 162 ggtggaggcg gtagcaaaaa cattattaat acttccatac ttaacctgcg ctacgagtct 60 aa 62 <210> 163 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM1/Forward primer <400> 163 tatgaatttc gtgccacggt aaagactgct gtt 33 <210> 164 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM1/Backward primer <400> 164 aacagcagtc tttaccgtgg cacgaaattc ata 33 <210> 165 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM2/Forward primer <400> 165 ttcgtcgtca ttacagaccc tcttgggccc tt 32 <210> 166 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM2/Backward primer <400> 166 aagggcccaa gagggtctgt aatgacgacg aa 32 <210> 167 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM3/Forward primer <400> 167 cacatgttt acgtccacgt atttgtttgg gtc 33 <210> 168 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> EGFP-RBD' SDM3/Backward primer <400> 168 gacccaaaca aaacgtggac gtaaacaatg tg 32 <210> 169 <211> 86 <212> DNA <213> Artificial Sequence <220> <223> EGFP-A2/Forward primer <400> 169 ggtggaggcg gtagcagagg gaatgtaatg acgacgaaca tataccttaa cagcagttga 60 gatccggctg ctaacaaagc ccgaaa 86 <210> 170 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> EGFP-A2/Backward primer <400> 170 gtggtggtgg tggtggtgct cg 22 <210> 171 <211> 75 <212> DNA <213> Artificial Sequence <220> <223> EGFP-Aurein1.2-A2/Forward primer <400> 171 gctaccgcct ccaccggcct gtttgacatc atcaaaaaaa tcgccgaaag ctttgctacc 60 gcctccaccg tggtg 75 <210> 172 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> EGFP-Aurein1.2-A2/Backward primer <400> 172 agagggaatg taatgacgac gaacatatac cttaacag 38 <210> 173 <211> 75 <212> DNA <213> Artificial Sequence <220> <223> EGFP-Aurein1.2-RBD/Forward primer <400> 173 gctaccgcct ccaccggcct gtttgacatc atcaaaaaaa tcgccgaaag ctttgctacc 60 gcctccaccg tggtg 75 <210> 174 <211> 44 <212> DNA <213> Artificial Sequence <220> <223> EGFP-Aurein1.2-RBD/Backward primer <400> 174 gactcgtagc gcaggttaag tatggaagta ttaataatgt tttt 44
Claims (28)
상기 신경-활성 물질은 보툴리눔 신경독소 경쇄인 것을 특징으로 하는, 신경세포 표적용 단백질.The method of claim 1,
The neuro-active substance is a botulinum neurotoxin light chain, characterized in that the neuron targeting protein.
상기 보툴리눔 신경독소 경쇄는 서열번호 1 내지 70으로 이루어진 군으로부터 선택되는 하나 이상의 아미노산 서열을 포함하는 것을 특징으로 하는, 신경세포 표적용 단백질.4. The method of claim 3,
The botulinum neurotoxin light chain is characterized in that it comprises one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 1 to 70, a protein for targeting nerve cells.
상기 신경-활성 물질은 저분자 신경-활성 물질인 것을 특징으로 하는, 신경세포 표적용 단백질.The method of claim 1,
The neuro-active substance is a small molecule neuro-active substance, characterized in that, a protein for targeting nerve cells.
상기 저분자 신경-활성 물질은 플라보노이드(flavonoid), 스테로이드(sterioid), 알칼로이드(alkaloid), 스틸베노이드(stilbenoid), 찰코노이드(chalconoid), 컬큐민(curcumin), 베라파밀(verapamil), 6-쇼가올(6-shogaol), 브로모데옥 시우리딘(bromodeoxyuridine), 5-플루오로우라신(5-fluorouracil), 요오드데옥시우리딘(iododeoxyuridine), 도파민(dopamine), 노르에피네프린(norepinephrine), 레티노산(retinoic acid), 파클리탁셀(paclitaxel), 카르보플라틴(carboplatin), 아이소나이아지드(isoniazid), 레보도파(levodopa), 및 카르무스틴(carmustine)으로 이루어진 군으로부터 선택된 하나 이상의 저분자 신경-활성 물질을 포함하는 것을 특징으로 하는, 신경세포 표적용 단백질.6. The method of claim 5,
The small molecule neuro-active substance is a flavonoid, steroid, alkaloid, stilbenoid, chalconoid, curcumin, verapamil, 6-shoga 6-shogaol, bromodeoxyuridine, 5-fluorouracil, iododeoxyuridine, dopamine, norepinephrine, retinoic acid ( retinoic acid), paclitaxel, carboplatin, isoniazid, levodopa, and carmustine comprising one or more small molecule neuro-active substances selected from the group consisting of Characterized in that, a protein for targeting nerve cells.
상기 신경-활성 물질은 저분자 신경-활성 물질을 함유하는 담체인 것을 특징으로 하는, 신경세포 표적용 단백질.The method of claim 1,
The neuronal-active substance is characterized in that the carrier containing a small molecule neuro-active substance, a protein for targeting nerve cells.
상기 담체는 유기미세입자, 무기미세입자 및 자가조립구조체(self-assembled structure)로 이루어진 군으로부터 선택된 하나 이상인 것을 특징으로 하는, 신경세포 표적용 단백질.8. The method of claim 7,
The carrier is at least one selected from the group consisting of organic microparticles, inorganic microparticles and self-assembled structures, a protein for nerve cell targeting.
상기 신경-활성 물질은 형광 단백질인 것을 특징으로 하는, 신경세포 표적용 단백질.The method of claim 1,
The neuronal-active substance is characterized in that the fluorescent protein, a protein for targeting nerve cells.
상기 형광 단백질은 서열번호 71 내지 75으로 이루어진 군으로부터 선택되는 하나 이상의 아미노산 서열을 포함하는 것을 특징으로 하는, 신경세포 표적용 단백질.10. The method of claim 9,
The fluorescent protein is characterized in that it comprises one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 71 to 75, a protein for targeting nerve cells.
상기 신경-활성 물질은 크산틴 유도체(xanthene derivatives), 시아닌 유도체(cyanine derivatives), 스쿠아레인 유도체(Squaraine derivatives), 스쿠아레인 로탁산 유도체(Squaraine Rotaxane derivatives), 나프탈렌 유도체(naphthalene derivatives), 쿠마린 유도체(coumarin derivatives), 옥사디아졸 유도체(oxadiazole derivatives), 안트라센 유도체(anthracene derivatives), 피렌 유도체(pyrene derivatives), 옥사진 유도체(oxazine derivatives), 아크리딘 유도체(acridine derivatives), 아릴메틴 유도체(arylmethine derivatives) 및 테트라피롤 유도체(tetrapyrrole derivatives)로 이루어진 군으로부터 선택된 하나 이상의 저분자 형광 화합물인 것을 특징으로 하는, 신경세포 표적용 단백질.The method of claim 1,
Said neuro-active substance is xanthene derivatives, cyanine derivatives, squaraine derivatives, squaraine rotaxane derivatives, naphthalene derivatives, coumarin Coumarin derivatives, oxadiazole derivatives, anthracene derivatives, pyrene derivatives, oxazine derivatives, acridine derivatives, arylmethine derivatives ( Arylmethine derivatives) and tetrapyrrole derivatives (tetrapyrrole derivatives) characterized in that at least one low-molecular-weight fluorescent compound selected from the group consisting of, a neuron targeting protein.
상기 신경-활성 물질은 저분자 형광 화합물을 함유하는 담체인 것을 특징으로 하는, 신경세포 표적용 단백질.The method of claim 1,
The neuron-active substance is a carrier containing a low molecular weight fluorescent compound, characterized in that the neuron targeting protein.
상기 담체는 유기미세입자, 무기미세입자 및 자가조립구조체(self-assembled structure)로 이루어진 군으로부터 선택된 어느 하나 이상인 것을 특징으로 하는, 신경세포 표적용 단백질.13. The method of claim 12,
The carrier is an organic microparticle, an inorganic microparticle, and a self-assembled structure (self-assembled structure) characterized in that any one or more selected from the group consisting of, a protein for nerve cell targeting.
상기 신경-활성 물질은 미세 반도체 입자인 것을 특징으로 하는, 신경세포 표적용 단백질.The method of claim 1,
The neuro-active material is characterized in that the fine semiconductor particles, nerve cell targeting protein.
상기 엔도소말 이스케이프 펩타이드는 서열번호 76 내지 98으로 이루어진 군으로부터 선택되는 하나 이상의 아미노산 서열을 포함하는 것을 특징으로 하는, 신경세포 표적용 단백질.The method of claim 1,
The endosomal escape peptide is characterized in that it comprises one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 76 to 98, nerve cell targeting protein.
상기 엔도소말 이스케이프 펩타이드는 하나 이상이 결합된 것을 특징으로 하는, 신경세포 표적용 단백질.The method of claim 1,
The endosomal escape peptide is characterized in that one or more is bound, a protein for a nerve cell target.
상기 보툴리눔 독소 단백질의 리셉터 바인딩 도메인은 서열번호 99 내지 115으로 이루어진 군으로부터 선택되는 하나 이상의 아미노산 서열을 포함하는 것을 특징으로 하는, 신경세포 표적용 단백질.The method of claim 1,
The receptor binding domain of the botulinum toxin protein is characterized in that it comprises one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 99 to 115, a protein for targeting nerve cells.
상기 보툴리눔 독소 타입A의 A2 펩타이드는 서열번호 116 내지 120으로 이루어진 군으로부터 선택되는 하나 이상의 아미노산 서열을 포함하는 것을 특징으로 하는, 신경세포 표적용 단백질.The method of claim 1,
The botulinum toxin type A A 2 peptide comprises at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 116 to 120, a protein for targeting nerve cells.
상기 신경-활성 물질은 신경병증 치료용 물질인 것을 특징으로 하는, 약학적 조성물.endosomal escape peptide to neuro-active material; and a receptor binding domain of a botulinum toxin protein or a neuronal targeting protein to which an A2 peptide of botulinum toxin type A is bound as an active ingredient. A therapeutic pharmaceutical composition comprising:
The neuro-active substance is a substance for the treatment of neuropathy, a pharmaceutical composition.
상기 신경병증 치료용 물질은 보툴리눔 신경독소 경쇄, 저분자 신경-활성 물질, 저분자 신경-활성 물질을 함유하는 담체 및 이의 조합으로 이루어진 군으로부터 선택된 어느 하나 이상인 것을 특징으로 하는, 약학적 조성물.20. The method of claim 19,
The substance for treating neuropathy is any one or more selected from the group consisting of a botulinum neurotoxin light chain, a low-molecular neurotoxin-active substance, a carrier containing a low-molecular neuroactive substance, and a combination thereof, a pharmaceutical composition.
상기 신경병증은 사시, 안검경련, 성대장애, 사경, 심근장애, 궤양, 위산역류질환, 식욕감소, 췌장질환, 튼 살, 절박성 요실금, 치열, 소아마비, 근육통, 엉덩이 기형, 다한증, 허리통증, 경부통, 만성두통 및 뇌신경장애로 이루어진 군으로부터 선택되는 하나 이상인 것을 특징으로 하는, 약학적 조성물.20. The method of claim 19,
The neuropathy is strabismus, blepharospasm, vocal cord disorder, torticollis, myocardial disorder, ulcer, acid reflux disease, decreased appetite, pancreatic disease, stretch marks, urge incontinence, dentition, polio, myalgia, hip deformity, hyperhidrosis, back pain, neck pain , A pharmaceutical composition, characterized in that at least one selected from the group consisting of chronic headaches and cranial nerve disorders.
상기 신경-활성 물질은 보툴리눔 신경독소 경쇄, 저분자 신경-활성 물질, 저분자 신경-활성 물질을 함유하는 담체, 및 이의 조합으로 이루어진 군으로부터 선택된 어느 하나 이상인 것을 특징으로 하는, 화장료 조성물.24. The method of claim 23,
The neuro-active substance is characterized in that at least one selected from the group consisting of a botulinum neurotoxin light chain, a small-molecule neuro-active substance, a carrier containing a low-molecular neuro-active substance, and a combination thereof, the cosmetic composition.
상기 신경-활성 물질은 뉴런을 선택적으로 발색시키는 것을 특징으로 하는, 신경계 뉴런 표시용 조성물.endosomal escape peptide to neuro-active material; and a receptor binding domain of a botulinum toxin protein or a neuron targeting protein to which an A2 peptide of botulinum toxin type A is bound as an active ingredient, a composition for displaying neurons in the nervous system as,
The neuro-active substance is characterized in that the neuron selectively develops color, a composition for displaying a neuron in the nervous system.
상기 신경-활성 물질은 형광 단백질, 저분자 형광 화합물, 저분자 형광 화합물을 함유하는 담체, 미세 반도체 입자, 및 이의 조합으로 이루어진 군으로부터 선택된 어느 하나 이상인 것을 특징으로 하는, 신경계 뉴런 표시용 조성물.27. The method of claim 26,
The neuro-active material is a fluorescent protein, a low-molecular-weight fluorescent compound, a carrier containing a low-molecular fluorescent compound, a micro semiconductor particle, and any one or more selected from the group consisting of a combination thereof, a composition for displaying neurons in the nervous system
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