                         SEQUENCE LISTING

<110>  SYNTAXIN LIMITED
 
<120>  CATIONIC NEUROTOXINS

<130>  P40989WO

<150>  GB 1312317.9
<151>  2013-07-09

<160>  10    

<170>  PatentIn version 3.5

<210>  1
<211>  3888
<212>  DNA
<213>  Clostridium botulinum

<400>  1
atgccattcg tcaacaagca attcaactac aaagacccag tcaacggcgt cgacatcgca       60

tacatcaaga ttccgaacgc cggtcaaatg cagccggtta aggcttttaa gatccacaac      120

aagatttggg ttatcccgga gcgtgacacc ttcacgaacc cggaagaagg cgatctgaac      180

ccgccaccgg aagcgaagca agtccctgtc agctactacg attcgacgta cctgagcacg      240

gataacgaaa aagataacta cctgaaaggt gtgaccaagc tgttcgaacg tatctacagc      300

acggatctgg gtcgcatgct gctgactagc attgttcgcg gtatcccgtt ctggggtggt      360

agcacgattg acaccgaact gaaggttatc gacactaact gcattaacgt tattcaaccg      420

gatggtagct atcgtagcga agagctgaat ctggtcatca ttggcccgag cgcagacatt      480

atccaattcg agtgcaagag ctttggtcac gaggttctga atctgacccg caatggctat      540

ggtagcaccc agtacattcg tttttcgccg gattttacct tcggctttga agagagcctg      600

gaggttgata ccaatccgtt gctgggtgcg ggcaaattcg ctaccgatcc ggctgtcacg      660

ctggcccatg aactgatcca cgcaggccac cgcctgtacg gcattgccat caacccaaac      720

cgtgtgttca aggttaatac gaatgcatac tacgagatga gcggcctgga agtcagcttc      780

gaagaactgc gcaccttcgg tggccatgac gctaaattca ttgacagctt gcaagagaat      840

gagttccgtc tgtactacta taacaaattc aaagacattg caagcacgtt gaacaaggcc      900

aaaagcatcg ttggtactac cgcgtcgttg cagtatatga agaatgtgtt taaagagaag      960

tacctgctgt ccgaggatac ctccggcaag tttagcgttg ataagctgaa gtttgacaaa     1020

ctgtacaaga tgctgaccga gatttacacc gaggacaact ttgtgaaatt cttcaaagtg     1080

ttgaatcgta aaacctatct gaattttgac aaagcggttt tcaagattaa catcgtgccg     1140

aaggtgaact acaccatcta tgacggtttt aacctgcgta acaccaacct ggcggcgaac     1200

tttaacggtc agaatacgga aatcaacaac atgaatttca cgaagttgaa gaacttcacg     1260

ggtctgttcg agttctataa gctgctgtgc gtgcgcggta tcatcaccag caaaaccaaa     1320

agcctggaca aaggctacaa caaggcgctg aatgacctgt gcattaaggt aaacaattgg     1380

gatctgttct tttcgccatc cgaagataat tttaccaacg acctgaacaa gggtgaagaa     1440

atcaccagcg atacgaatat tgaagcagcg gaagagaata tcagcctgga tctgatccag     1500

cagtactatc tgacctttaa cttcgacaat gaaccggaga acattagcat tgagaatctg     1560

agcagcgaca ttatcggtca gctggaactg atgccgaata tcgaacgttt cccgaacggc     1620

aaaaagtacg agctggacaa gtacactatg ttccattacc tgcgtgcaca ggagtttgaa     1680

cacggtaaaa gccgtatcgc gctgaccaac agcgttaacg aggccctgct gaacccgagc     1740

cgtgtctata ccttcttcag cagcgactat gttaagaaag tgaacaaagc cactgaggcc     1800

gcgatgttcc tgggctgggt ggaacagctg gtatatgact tcacggacga gacgagcgaa     1860

gtgagcacta ccgacaaaat tgctgatatt accatcatta tcccgtatat tggtccggca     1920

ctgaacattg gcaacatgct gtacaaagac gattttgtgg gtgccctgat cttctccggt     1980

gccgtgattc tgctggagtt cattccggag attgcgatcc cggtgttggg taccttcgcg     2040

ctggtgtcct acatcgcgaa taaggttctg acggttcaga ccatcgataa cgcgctgtcg     2100

aaacgtaatg aaaaatggga cgaggtttac aaatacattg ttacgaattg gctggcgaaa     2160

gtcaataccc agatcgacct gatccgtaag aaaatgaaag aggcgctgga gaatcaggcg     2220

gaggccacca aagcaattat caactaccaa tacaaccagt acacggaaga agagaagaat     2280

aacattaact tcaatatcga tgatttgagc agcaagctga atgaatctat caacaaagcg     2340

atgatcaata tcaacaagtt tttgaatcag tgtagcgttt cgtacctgat gaatagcatg     2400

attccgtatg gcgtcaaacg tctggaggac ttcgacgcca gcctgaaaga tgcgttgctg     2460

aaatacattt acgacaatcg tggtacgctg attggccaag ttgaccgctt gaaagacaaa     2520

gttaacaata ccctgagcac cgacatccca tttcaactga gcaagtatgt tgataatcaa     2580

cgtctgttga gcactttcac cgagtatatc aaaaacatca tcaatactag cattctgaac     2640

ctgcgttacg agagcaatca tctgattgat ctgagccgtt atgcaagcaa gatcaacatc     2700

ggtagcaagg tcaattttga cccgatcgat aagaaccaga tccagctgtt taatctggaa     2760

tcgagcaaaa ttgaggttat cctgaaaaac gccattgtct acaactccat gtacgagaat     2820

ttctccacca gcttctggat tcgcatcccg aaatacttca acagcattag cctgaacaac     2880

gagtatacta tcatcaactg tatggagaac aacagcggtt ggaaggtgtc tctgaactat     2940

ggtgagatca tttggacctt gcaggacacc caagagatca agcagcgcgt cgtgttcaag     3000

tactctcaaa tgatcaacat ttccgattac attaatcgtt ggatcttcgt gaccattacg     3060

aataaccgtc tgaataacag caagatttac atcaatggtc gcttgatcga tcagaaaccg     3120

attagcaacc tgggtaatat ccacgcaagc aacaacatta tgttcaaatt ggacggttgc     3180

cgcgataccc atcgttatat ctggatcaag tatttcaacc tgtttgataa agaactgaat     3240

gagaaggaga tcaaagattt gtatgacaac caatctaaca gcggcatttt gaaggacttc     3300

tggggcgatt atctgcaata cgataagccg tactatatgc tgaacctgta tgatccgaac     3360

aaatatgtgg atgtcaataa tgtgggtatt cgtggttaca tgtatttgaa gggtccgcgt     3420

ggcagcgtta tgacgaccaa catttacctg aactctagcc tgtaccgtgg tacgaaattc     3480

atcattaaga aatatgccag cggcaacaaa gataacattg tgcgtaataa cgatcgtgtc     3540

tacatcaacg tggtcgtgaa gaataaagag taccgtctgg cgaccaacgc ttcgcaggcg     3600

ggtgttgaga aaattctgag cgcgttggag atccctgatg tcggtaatct gagccaagtc     3660

gtggttatga agagcaagaa cgaccagggt atcactaaca agtgcaagat gaacctgcaa     3720

gacaacaatg gtaacgacat cggctttatt ggtttccacc agttcaacaa tattgctaaa     3780

ctggtagcga gcaattggta caatcgtcag attgagcgca gcagccgtac tttgggctgt     3840

agctgggagt ttatcccggt cgatgatggt tggggcgaac gtccgctg                  3888


<210>  2
<211>  1296
<212>  PRT
<213>  Clostridium botulinum

<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 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             


Ala Leu Asn Asp Leu Cys Ile Lys Val Asn Asn Trp Asp Leu Phe Phe 
    450                 455                 460                 


Ser Pro Ser Glu Asp Asn Phe Thr Asn Asp Leu Asn Lys Gly Glu Glu 
465                 470                 475                 480 


Ile Thr Ser Asp Thr Asn Ile Glu Ala Ala Glu Glu Asn Ile Ser Leu 
                485                 490                 495     


Asp Leu Ile Gln Gln Tyr Tyr Leu Thr Phe Asn Phe Asp Asn Glu Pro 
            500                 505                 510         


Glu Asn Ile Ser Ile Glu Asn Leu Ser Ser Asp Ile Ile Gly Gln Leu 
        515                 520                 525             


Glu Leu Met Pro Asn Ile Glu Arg Phe Pro Asn Gly Lys Lys Tyr Glu 
    530                 535                 540                 


Leu Asp Lys Tyr Thr Met Phe His Tyr Leu Arg Ala Gln Glu Phe Glu 
545                 550                 555                 560 


His Gly Lys Ser Arg Ile Ala Leu Thr Asn Ser Val Asn Glu Ala Leu 
                565                 570                 575     


Leu Asn Pro Ser Arg Val Tyr Thr Phe Phe Ser Ser Asp Tyr Val Lys 
            580                 585                 590         


Lys Val Asn Lys Ala Thr Glu Ala Ala Met Phe Leu Gly Trp Val Glu 
        595                 600                 605             


Gln Leu Val Tyr Asp Phe Thr Asp Glu Thr Ser Glu Val Ser Thr Thr 
    610                 615                 620                 


Asp Lys Ile Ala Asp Ile Thr Ile Ile Ile Pro Tyr Ile Gly Pro Ala 
625                 630                 635                 640 


Leu Asn Ile Gly Asn Met Leu Tyr Lys Asp Asp Phe Val Gly Ala Leu 
                645                 650                 655     


Ile Phe Ser Gly Ala Val Ile Leu Leu Glu Phe Ile Pro Glu Ile Ala 
            660                 665                 670         


Ile Pro Val Leu Gly Thr Phe Ala Leu Val Ser Tyr Ile Ala Asn Lys 
        675                 680                 685             


Val Leu Thr Val Gln Thr Ile Asp Asn Ala Leu Ser Lys Arg Asn Glu 
    690                 695                 700                 


Lys Trp Asp Glu Val Tyr Lys Tyr Ile Val Thr Asn Trp Leu Ala Lys 
705                 710                 715                 720 


Val Asn Thr Gln Ile Asp Leu Ile Arg Lys Lys Met Lys Glu Ala Leu 
                725                 730                 735     


Glu Asn Gln Ala Glu Ala Thr Lys Ala Ile Ile Asn Tyr Gln Tyr Asn 
            740                 745                 750         


Gln Tyr Thr Glu Glu Glu Lys Asn Asn Ile Asn Phe Asn Ile Asp Asp 
        755                 760                 765             


Leu Ser Ser Lys Leu Asn Glu Ser Ile Asn Lys Ala Met Ile Asn Ile 
    770                 775                 780                 


Asn Lys Phe Leu Asn Gln Cys Ser Val Ser Tyr Leu Met Asn Ser Met 
785                 790                 795                 800 


Ile Pro Tyr Gly Val Lys Arg Leu Glu Asp Phe Asp Ala Ser Leu Lys 
                805                 810                 815     


Asp Ala Leu Leu Lys Tyr Ile Tyr Asp Asn Arg Gly Thr Leu Ile Gly 
            820                 825                 830         


Gln Val Asp Arg Leu Lys Asp Lys Val Asn Asn Thr Leu Ser Thr Asp 
        835                 840                 845             


Ile Pro Phe Gln Leu Ser Lys Tyr Val Asp Asn Gln Arg Leu Leu Ser 
    850                 855                 860                 


Thr Phe Thr Glu Tyr Ile Lys Asn Ile Ile Asn Thr Ser Ile Leu Asn 
865                 870                 875                 880 


Leu Arg Tyr Glu Ser Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser 
                885                 890                 895     


Lys Ile Asn Ile Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn 
            900                 905                 910         


Gln Ile Gln Leu Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu 
        915                 920                 925             


Lys Asn Ala Ile Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser 
    930                 935                 940                 


Phe Trp Ile Arg Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn 
945                 950                 955                 960 


Glu Tyr Thr Ile Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val 
                965                 970                 975     


Ser Leu Asn Tyr Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu 
            980                 985                 990         


Ile Lys Gln Arg Val Val Phe Lys  Tyr Ser Gln Met Ile  Asn Ile Ser 
        995                 1000                 1005             


Asp Tyr  Ile Asn Arg Trp Ile  Phe Val Thr Ile Thr  Asn Asn Arg 
    1010                 1015                 1020             


Leu Asn  Asn Ser Lys Ile Tyr  Ile Asn Gly Arg Leu  Ile Asp Gln 
    1025                 1030                 1035             


Lys Pro  Ile Ser Asn Leu Gly  Asn Ile His Ala Ser  Asn Asn Ile 
    1040                 1045                 1050             


Met Phe  Lys Leu Asp Gly Cys  Arg Asp Thr His Arg  Tyr Ile Trp 
    1055                 1060                 1065             


Ile Lys  Tyr Phe Asn Leu Phe  Asp Lys Glu Leu Asn  Glu Lys Glu 
    1070                 1075                 1080             


Ile Lys  Asp Leu Tyr Asp Asn  Gln Ser Asn Ser Gly  Ile Leu Lys 
    1085                 1090                 1095             


Asp Phe  Trp Gly Asp Tyr Leu  Gln Tyr Asp Lys Pro  Tyr Tyr Met 
    1100                 1105                 1110             


Leu Asn  Leu Tyr Asp Pro Asn  Lys Tyr Val Asp Val  Asn Asn Val 
    1115                 1120                 1125             


Gly Ile  Arg Gly Tyr Met Tyr  Leu Lys Gly Pro Arg  Gly Ser Val 
    1130                 1135                 1140             


Met Thr  Thr Asn Ile Tyr Leu  Asn Ser Ser Leu Tyr  Arg Gly Thr 
    1145                 1150                 1155             


Lys Phe  Ile Ile Lys Lys Tyr  Ala Ser Gly Asn Lys  Asp Asn Ile 
    1160                 1165                 1170             


Val Arg  Asn Asn Asp Arg Val  Tyr Ile Asn Val Val  Val Lys Asn 
    1175                 1180                 1185             


Lys Glu  Tyr Arg Leu Ala Thr  Asn Ala Ser Gln Ala  Gly Val Glu 
    1190                 1195                 1200             


Lys Ile  Leu Ser Ala Leu Glu  Ile Pro Asp Val Gly  Asn Leu Ser 
    1205                 1210                 1215             


Gln Val  Val Val Met Lys Ser  Lys Asn Asp Gln Gly  Ile Thr Asn 
    1220                 1225                 1230             


Lys Cys  Lys Met Asn Leu Gln  Asp Asn Asn Gly Asn  Asp Ile Gly 
    1235                 1240                 1245             


Phe Ile  Gly Phe His Gln Phe  Asn Asn Ile Ala Lys  Leu Val Ala 
    1250                 1255                 1260             


Ser Asn  Trp Tyr Asn Arg Gln  Ile Glu Arg Ser Ser  Arg Thr Leu 
    1265                 1270                 1275             


Gly Cys  Ser Trp Glu Phe Ile  Pro Val Asp Asp Gly  Trp Gly Glu 
    1280                 1285                 1290             


Arg Pro  Leu 
    1295     


<210>  3
<211>  3888
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Engineered BoNT/A1

<400>  3
atgccattcg tcaacaagca attcaactac aaagacccag tcaacggcgt cgacatcgca       60

tacatcaaga ttccgaacgc cggtcaaatg cagccggtta aggcttttaa gatccacaac      120

aagatttggg ttatcccgga gcgtgacacc ttcacgaacc cggaagaagg cgatctgaac      180

ccgccaccgg aagcgaagca agtccctgtc agctactacg attcgacgta cctgagcacg      240

gataacgaaa aagataacta cctgaaaggt gtgaccaagc tgttcgaacg tatctacagc      300

acggatctgg gtcgcatgct gctgactagc attgttcgcg gtatcccgtt ctggggtggt      360

agcacgattg acaccgaact gaaggttatc gacactaact gcattaacgt tattcaaccg      420

gatggtagct atcgtagcga agagctgaat ctggtcatca ttggcccgag cgcagacatt      480

atccaattcg agtgcaagag ctttggtcac gaggttctga atctgacccg caatggctat      540

ggtagcaccc agtacattcg tttttcgccg gattttacct tcggctttga agagagcctg      600

gaggttgata ccaatccgtt gctgggtgcg ggcaaattcg ctaccgatcc ggctgtcacg      660

ctggcccatg aactgatcca cgcaggccac cgcctgtacg gcattgccat caacccaaac      720

cgtgtgttca aggttaatac gaatgcatac tacgagatga gcggcctgga agtcagcttc      780

gaagaactgc gcaccttcgg tggccatgac gctaaattca ttgacagctt gcaagagaat      840

gagttccgtc tgtactacta taacaaattc aaagacattg caagcacgtt gaacaaggcc      900

aaaagcatcg ttggtactac cgcgtcgttg cagtatatga agaatgtgtt taaagagaag      960

tacctgctgt ccgaggatac ctccggcaag tttagcgttg ataagctgaa gtttgacaaa     1020

ctgtacaaga tgctgaccga gatttacacc gaggacaact ttgtgaaatt cttcaaagtg     1080

ttgaatcgta aaacctatct gaattttgac aaagcggttt tcaagattaa catcgtgccg     1140

aaggtgaact acaccatcta tgacggtttt aacctgcgta acaccaacct ggcggcgaac     1200

tttaacggtc agaatacgga aatcaacaac atgaatttca cgaagttgaa gaacttcacg     1260

ggtctgttcg agttctataa gctgctgtgc gtgcgcggta tcatcaccag caaaaccaaa     1320

agcctggaca aaggctacaa caaggcgctg aatgacctgt gcattaaggt aaacaattgg     1380

gatctgttct tttcgccatc cgaagataat tttaccaacg acctgaacaa gggtgaagaa     1440

atcaccagcg atacgaatat tgaagcagcg gaagagaata tcagcctgga tctgatccag     1500

cagtactatc tgacctttaa cttcgacaat gaaccggaga acattagcat tgagaatctg     1560

agcagcgaca ttatcggtca gctggaactg atgccgaata tcgaacgttt cccgaacggc     1620

aaaaagtacg agctggacaa gtacactatg ttccattacc tgcgtgcaca ggagtttgaa     1680

cacggtaaaa gccgtatcgc gctgaccaac agcgttaacg aggccctgct gaacccgagc     1740

cgtgtctata ccttcttcag cagcgactat gttaagaaag tgaacaaagc cactgaggcc     1800

gcgatgttcc tgggctgggt ggaacagctg gtatatgact tcacggacga gacgagcgaa     1860

gtgagcacta ccgacaaaat tgctgatatt accatcatta tcccgtatat tggtccggca     1920

ctgaacattg gcaacatgct gtacaaagac gattttgtgg gtgccctgat cttctccggt     1980

gccgtgattc tgctggagtt cattccggag attgcgatcc cggtgttggg taccttcgcg     2040

ctggtgtcct acatcgcgaa taaggttctg acggttcaga ccatcgataa cgcgctgtcg     2100

aaacgtaatg aaaaatggga cgaggtttac aaatacattg ttacgaattg gctggcgaaa     2160

gtcaataccc agatcgacct gatccgtaag aaaatgaaag aggcgctgga gaatcaggcg     2220

gaggccacca aagcaattat caactaccaa tacaaccagt acacggaaga agagaagaat     2280

aacattaact tcaatatcga tgatttgagc agcaagctga atgaatctat caacaaagcg     2340

atgatcaata tcaacaagtt tttgaatcag tgtagcgttt cgtacctgat gaatagcatg     2400

attccgtatg gcgtcaaacg tctggaggac ttcgacgcca gcctgaaaga tgcgttgctg     2460

aaatacattt acgacaatcg tggtacgctg attggccaag ttgaccgctt gaaagacaaa     2520

gttaacaata ccctgagcac cgacatccca tttcaactga gcaagtatgt tgataatcaa     2580

cgtctgttga gcactttcac cgagtatatc aaaaacatca tcaatactag cattctgaac     2640

ctgcgttacg agagcaagca tctgattgat ctgagccgtt atgctagcaa gatcaacatc     2700

ggtagcaagg tcaattttga cccgatcgat aagaaccaga tccagctgtt taatctggaa     2760

tcgagcaaaa ttgaggttat cctgaaaaag gccattgtct acaactccat gtacgagaat     2820

ttctccacca gcttctggat tcgcatcccg aaatacttca acaagattag cctgaacaac     2880

gagtatacta tcatcaactg tatggagaac aacagcggtt ggaaggtgtc tctgaactat     2940

ggtgagatca tttggacctt gcaggacacc aaagagatca agcagcgcgt cgtgttcaag     3000

tactctcaaa tgatcaacat ttccgattac attaatcgtt ggatcttcgt gaccattacg     3060

aataaccgtc tgaataagag caagatttac atcaatggtc gcttgatcga tcagaaaccg     3120

attagcaacc tgggtaatat ccacgcaagc aacaagatta tgttcaaatt ggacggttgc     3180

cgcgataccc atcgttatat ctggatcaag tatttcaacc tgtttgataa agaactgaat     3240

gagaaggaga tcaaagattt gtatgacaac caatctaaca gcggcatttt gaaggacttc     3300

tggggcgatt atctgcaata cgataagccg tactatatgc tgaacctgta tgatccgaac     3360

aaatatgtgg atgtcaataa tgtgggtatt cgtggttaca tgtatttgaa gggtccgcgt     3420

ggcagcgtta tgacgaccaa catttacctg aactctagcc tgtaccgtgg tacgaaattc     3480

atcattaaga aatatgccag cggcaacaaa gataacattg tgcgtaataa cgatcgtgtc     3540

tacatcaacg tggtcgtgaa gaataaagag taccgtctgg cgaccaacgc ttcgcaggcg     3600

ggtgttgaga aaattctgag cgcgttggag atccctgatg tcggtaatct gagccaagtc     3660

gtggttatga agagcaagaa cgacaagggt atcactaaca agtgcaagat gaacctgcaa     3720

gacaacaatg gtaacgacat cggctttatt ggtttccacc agttcaacaa tattgctaaa     3780

ctggtagcga gcaattggta caatcgtcag attgagcgca gcagccgtac tttgggctgt     3840

agctgggagt ttatcccggt cgatgatggt tggggcgaac gtccgctg                  3888


<210>  4
<211>  1296
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Engineered BoNT/A1

<400>  4

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             


Ala Leu Asn Asp Leu Cys Ile Lys Val Asn Asn Trp Asp Leu Phe Phe 
    450                 455                 460                 


Ser Pro Ser Glu Asp Asn Phe Thr Asn Asp Leu Asn Lys Gly Glu Glu 
465                 470                 475                 480 


Ile Thr Ser Asp Thr Asn Ile Glu Ala Ala Glu Glu Asn Ile Ser Leu 
                485                 490                 495     


Asp Leu Ile Gln Gln Tyr Tyr Leu Thr Phe Asn Phe Asp Asn Glu Pro 
            500                 505                 510         


Glu Asn Ile Ser Ile Glu Asn Leu Ser Ser Asp Ile Ile Gly Gln Leu 
        515                 520                 525             


Glu Leu Met Pro Asn Ile Glu Arg Phe Pro Asn Gly Lys Lys Tyr Glu 
    530                 535                 540                 


Leu Asp Lys Tyr Thr Met Phe His Tyr Leu Arg Ala Gln Glu Phe Glu 
545                 550                 555                 560 


His Gly Lys Ser Arg Ile Ala Leu Thr Asn Ser Val Asn Glu Ala Leu 
                565                 570                 575     


Leu Asn Pro Ser Arg Val Tyr Thr Phe Phe Ser Ser Asp Tyr Val Lys 
            580                 585                 590         


Lys Val Asn Lys Ala Thr Glu Ala Ala Met Phe Leu Gly Trp Val Glu 
        595                 600                 605             


Gln Leu Val Tyr Asp Phe Thr Asp Glu Thr Ser Glu Val Ser Thr Thr 
    610                 615                 620                 


Asp Lys Ile Ala Asp Ile Thr Ile Ile Ile Pro Tyr Ile Gly Pro Ala 
625                 630                 635                 640 


Leu Asn Ile Gly Asn Met Leu Tyr Lys Asp Asp Phe Val Gly Ala Leu 
                645                 650                 655     


Ile Phe Ser Gly Ala Val Ile Leu Leu Glu Phe Ile Pro Glu Ile Ala 
            660                 665                 670         


Ile Pro Val Leu Gly Thr Phe Ala Leu Val Ser Tyr Ile Ala Asn Lys 
        675                 680                 685             


Val Leu Thr Val Gln Thr Ile Asp Asn Ala Leu Ser Lys Arg Asn Glu 
    690                 695                 700                 


Lys Trp Asp Glu Val Tyr Lys Tyr Ile Val Thr Asn Trp Leu Ala Lys 
705                 710                 715                 720 


Val Asn Thr Gln Ile Asp Leu Ile Arg Lys Lys Met Lys Glu Ala Leu 
                725                 730                 735     


Glu Asn Gln Ala Glu Ala Thr Lys Ala Ile Ile Asn Tyr Gln Tyr Asn 
            740                 745                 750         


Gln Tyr Thr Glu Glu Glu Lys Asn Asn Ile Asn Phe Asn Ile Asp Asp 
        755                 760                 765             


Leu Ser Ser Lys Leu Asn Glu Ser Ile Asn Lys Ala Met Ile Asn Ile 
    770                 775                 780                 


Asn Lys Phe Leu Asn Gln Cys Ser Val Ser Tyr Leu Met Asn Ser Met 
785                 790                 795                 800 


Ile Pro Tyr Gly Val Lys Arg Leu Glu Asp Phe Asp Ala Ser Leu Lys 
                805                 810                 815     


Asp Ala Leu Leu Lys Tyr Ile Tyr Asp Asn Arg Gly Thr Leu Ile Gly 
            820                 825                 830         


Gln Val Asp Arg Leu Lys Asp Lys Val Asn Asn Thr Leu Ser Thr Asp 
        835                 840                 845             


Ile Pro Phe Gln Leu Ser Lys Tyr Val Asp Asn Gln Arg Leu Leu Ser 
    850                 855                 860                 


Thr Phe Thr Glu Tyr Ile Lys Asn Ile Ile Asn Thr Ser Ile Leu Asn 
865                 870                 875                 880 


Leu Arg Tyr Glu Ser Lys His Leu Ile Asp Leu Ser Arg Tyr Ala Ser 
                885                 890                 895     


Lys Ile Asn Ile Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn 
            900                 905                 910         


Gln Ile Gln Leu Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu 
        915                 920                 925             


Lys Lys Ala Ile Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser 
    930                 935                 940                 


Phe Trp Ile Arg Ile Pro Lys Tyr Phe Asn Lys Ile Ser Leu Asn Asn 
945                 950                 955                 960 


Glu Tyr Thr Ile Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val 
                965                 970                 975     


Ser Leu Asn Tyr Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Lys Glu 
            980                 985                 990         


Ile Lys Gln Arg Val Val Phe Lys  Tyr Ser Gln Met Ile  Asn Ile Ser 
        995                 1000                 1005             


Asp Tyr  Ile Asn Arg Trp Ile  Phe Val Thr Ile Thr  Asn Asn Arg 
    1010                 1015                 1020             


Leu Asn  Lys Ser Lys Ile Tyr  Ile Asn Gly Arg Leu  Ile Asp Gln 
    1025                 1030                 1035             


Lys Pro  Ile Ser Asn Leu Gly  Asn Ile His Ala Ser  Asn Lys Ile 
    1040                 1045                 1050             


Met Phe  Lys Leu Asp Gly Cys  Arg Asp Thr His Arg  Tyr Ile Trp 
    1055                 1060                 1065             


Ile Lys  Tyr Phe Asn Leu Phe  Asp Lys Glu Leu Asn  Glu Lys Glu 
    1070                 1075                 1080             


Ile Lys  Asp Leu Tyr Asp Asn  Gln Ser Asn Ser Gly  Ile Leu Lys 
    1085                 1090                 1095             


Asp Phe  Trp Gly Asp Tyr Leu  Gln Tyr Asp Lys Pro  Tyr Tyr Met 
    1100                 1105                 1110             


Leu Asn  Leu Tyr Asp Pro Asn  Lys Tyr Val Asp Val  Asn Asn Val 
    1115                 1120                 1125             


Gly Ile  Arg Gly Tyr Met Tyr  Leu Lys Gly Pro Arg  Gly Ser Val 
    1130                 1135                 1140             


Met Thr  Thr Asn Ile Tyr Leu  Asn Ser Ser Leu Tyr  Arg Gly Thr 
    1145                 1150                 1155             


Lys Phe  Ile Ile Lys Lys Tyr  Ala Ser Gly Asn Lys  Asp Asn Ile 
    1160                 1165                 1170             


Val Arg  Asn Asn Asp Arg Val  Tyr Ile Asn Val Val  Val Lys Asn 
    1175                 1180                 1185             


Lys Glu  Tyr Arg Leu Ala Thr  Asn Ala Ser Gln Ala  Gly Val Glu 
    1190                 1195                 1200             


Lys Ile  Leu Ser Ala Leu Glu  Ile Pro Asp Val Gly  Asn Leu Ser 
    1205                 1210                 1215             


Gln Val  Val Val Met Lys Ser  Lys Asn Asp Lys Gly  Ile Thr Asn 
    1220                 1225                 1230             


Lys Cys  Lys Met Asn Leu Gln  Asp Asn Asn Gly Asn  Asp Ile Gly 
    1235                 1240                 1245             


Phe Ile  Gly Phe His Gln Phe  Asn Asn Ile Ala Lys  Leu Val Ala 
    1250                 1255                 1260             


Ser Asn  Trp Tyr Asn Arg Gln  Ile Glu Arg Ser Ser  Arg Thr Leu 
    1265                 1270                 1275             


Gly Cys  Ser Trp Glu Phe Ile  Pro Val Asp Asp Gly  Trp Gly Glu 
    1280                 1285                 1290             


Arg Pro  Leu 
    1295     


<210>  5
<211>  3888
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Engineered BoNT/A1

<400>  5
atgccattcg tcaacaagca attcaactac aaagacccag tcaacggcgt cgacatcgca       60

tacatcaaga ttccgaacgc cggtcaaatg cagccggtta aggcttttaa gatccacaac      120

aagatttggg ttatcccgga gcgtgacacc ttcacgaacc cggaagaagg cgatctgaac      180

ccgccaccgg aagcgaagca agtccctgtc agctactacg attcgacgta cctgagcacg      240

gataacgaaa aagataacta cctgaaaggt gtgaccaagc tgttcgaacg tatctacagc      300

acggatctgg gtcgcatgct gctgactagc attgttcgcg gtatcccgtt ctggggtggt      360

agcacgattg acaccgaact gaaggttatc gacactaact gcattaacgt tattcaaccg      420

gatggtagct atcgtagcga agagctgaat ctggtcatca ttggcccgag cgcagacatt      480

atccaattcg agtgcaagag ctttggtcac gaggttctga atctgacccg caatggctat      540

ggtagcaccc agtacattcg tttttcgccg gattttacct tcggctttga agagagcctg      600

gaggttgata ccaatccgtt gctgggtgcg ggcaaattcg ctaccgatcc ggctgtcacg      660

ctggcccatg aactgatcca cgcaggccac cgcctgtacg gcattgccat caacccaaac      720

cgtgtgttca aggttaatac gaatgcatac tacgagatga gcggcctgga agtcagcttc      780

gaagaactgc gcaccttcgg tggccatgac gctaaattca ttgacagctt gcaagagaat      840

gagttccgtc tgtactacta taacaaattc aaagacattg caagcacgtt gaacaaggcc      900

aaaagcatcg ttggtactac cgcgtcgttg cagtatatga agaatgtgtt taaagagaag      960

tacctgctgt ccgaggatac ctccggcaag tttagcgttg ataagctgaa gtttgacaaa     1020

ctgtacaaga tgctgaccga gatttacacc gaggacaact ttgtgaaatt cttcaaagtg     1080

ttgaatcgta aaacctatct gaattttgac aaagcggttt tcaagattaa catcgtgccg     1140

aaggtgaact acaccatcta tgacggtttt aacctgcgta acaccaacct ggcggcgaac     1200

tttaacggtc agaatacgga aatcaacaac atgaatttca cgaagttgaa gaacttcacg     1260

ggtctgttcg agttctataa gctgctgtgc gtgcgcggta tcatcaccag caaaaccaaa     1320

agcctggaca aaggctacaa caaggcgctg aatgacctgt gcattaaggt aaacaattgg     1380

gatctgttct tttcgccatc cgaagataat tttaccaacg acctgaacaa gggtgaagaa     1440

atcaccagcg atacgaatat tgaagcagcg gaagagaata tcagcctgga tctgatccag     1500

cagtactatc tgacctttaa cttcgacaat gaaccggaga acattagcat tgagaatctg     1560

agcagcgaca ttatcggtca gctggaactg atgccgaata tcgaacgttt cccgaacggc     1620

aaaaagtacg agctggacaa gtacactatg ttccattacc tgcgtgcaca ggagtttgaa     1680

cacggtaaaa gccgtatcgc gctgaccaac agcgttaacg aggccctgct gaacccgagc     1740

cgtgtctata ccttcttcag cagcgactat gttaagaaag tgaacaaagc cactgaggcc     1800

gcgatgttcc tgggctgggt ggaacagctg gtatatgact tcacggacga gacgagcgaa     1860

gtgagcacta ccgacaaaat tgctgatatt accatcatta tcccgtatat tggtccggca     1920

ctgaacattg gcaacatgct gtacaaagac gattttgtgg gtgccctgat cttctccggt     1980

gccgtgattc tgctggagtt cattccggag attgcgatcc cggtgttggg taccttcgcg     2040

ctggtgtcct acatcgcgaa taaggttctg acggttcaga ccatcgataa cgcgctgtcg     2100

aaacgtaatg aaaaatggga cgaggtttac aaatacattg ttacgaattg gctggcgaaa     2160

gtcaataccc agatcgacct gatccgtaag aaaatgaaag aggcgctgga gaatcaggcg     2220

gaggccacca aagcaattat caactaccaa tacaaccagt acacggaaga agagaagaat     2280

aacattaact tcaatatcga tgatttgagc agcaagctga atgaatctat caacaaagcg     2340

atgatcaata tcaacaagtt tttgaatcag tgtagcgttt cgtacctgat gaatagcatg     2400

attccgtatg gcgtcaaacg tctggaggac ttcgacgcca gcctgaaaga tgcgttgctg     2460

aaatacattt acgacaatcg tggtacgctg attggccaag ttgaccgctt gaaagacaaa     2520

gttaacaata ccctgagcac cgacatccca tttcaactga gcaagtatgt tgataatcaa     2580

cgtctgttga gcactttcac cgagtatatc aaaaacatca tcaatactag cattctgaac     2640

ctgcgttacg agagcaatca tctgattgat ctgagccgtt atgctagcaa gatcaacatc     2700

ggtagcaagg tcaattttga cccgatcgat aagaaccaga tccagctgtt taatctggaa     2760

tcgagcaaaa ttgaggttat cctgaaaaag gccattgtct acaactccat gtacgagaat     2820

ttctccacca gcttctggat tcgcatcccg aaatacttca agaagattag cctgaacaac     2880

gagtatacta tcatcaactg tatggagaac aacagcggtt ggaaggtgtc tctgaactat     2940

ggtgagatca tttggacctt gcaggacacc aaagagatca agcagcgcgt cgtgttcaag     3000

tactctcaaa tgatcaacat ttccgattac attaatcgtt ggatcttcgt gaccattacg     3060

aataaccgtc tgaataagag caagatttac atcaatggtc gcttgatcga tcagaaaccg     3120

attagcaacc tgggtaatat ccacgcaagc aacaagatta tgttcaaatt ggacggttgc     3180

cgcgataccc atcgttatat ctggatcaag tatttcaacc tgtttgataa agaactgaat     3240

gagaaggaga tcaaagattt gtatgacaac caatctaaca gcggcatttt gaaggacttc     3300

tggggcgatt atctgcaata cgataagccg tactatatgc tgaacctgta tgatccgaac     3360

aaatatgtgg atgtcaataa tgtgggtatt cgtggttaca tgtatttgaa gggtccgcgt     3420

ggcagcgtta tgacgaccaa catttacctg aactctagcc tgtaccgtgg tacgaaattc     3480

atcattaaga aatatgccag cggcaacaaa gataacattg tgcgtaataa cgatcgtgtc     3540

tacatcaacg tggtcgtgaa gaataaagag taccgtctgg cgaccaacgc ttcgcaggcg     3600

ggtgttgaga aaattctgag cgcgttggag atccctgatg tcggtaatct gagccaagtc     3660

gtggttatga agagcaagaa cgacaagggt atcactaaca agtgcaagat gaacctgcaa     3720

gacaacaatg gtaacgacat cggctttatt ggtttccacc agttcaacaa tattgctaaa     3780

ctggtagcga gcaattggta caatcgtcag attgagcgca gcagccgtac tttgggctgt     3840

agctgggagt ttatcccggt cgatgatggt tggggcgaac gtccgctg                  3888


<210>  6
<211>  1296
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Engineered BoNT/A1

<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 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             


Ala Leu Asn Asp Leu Cys Ile Lys Val Asn Asn Trp Asp Leu Phe Phe 
    450                 455                 460                 


Ser Pro Ser Glu Asp Asn Phe Thr Asn Asp Leu Asn Lys Gly Glu Glu 
465                 470                 475                 480 


Ile Thr Ser Asp Thr Asn Ile Glu Ala Ala Glu Glu Asn Ile Ser Leu 
                485                 490                 495     


Asp Leu Ile Gln Gln Tyr Tyr Leu Thr Phe Asn Phe Asp Asn Glu Pro 
            500                 505                 510         


Glu Asn Ile Ser Ile Glu Asn Leu Ser Ser Asp Ile Ile Gly Gln Leu 
        515                 520                 525             


Glu Leu Met Pro Asn Ile Glu Arg Phe Pro Asn Gly Lys Lys Tyr Glu 
    530                 535                 540                 


Leu Asp Lys Tyr Thr Met Phe His Tyr Leu Arg Ala Gln Glu Phe Glu 
545                 550                 555                 560 


His Gly Lys Ser Arg Ile Ala Leu Thr Asn Ser Val Asn Glu Ala Leu 
                565                 570                 575     


Leu Asn Pro Ser Arg Val Tyr Thr Phe Phe Ser Ser Asp Tyr Val Lys 
            580                 585                 590         


Lys Val Asn Lys Ala Thr Glu Ala Ala Met Phe Leu Gly Trp Val Glu 
        595                 600                 605             


Gln Leu Val Tyr Asp Phe Thr Asp Glu Thr Ser Glu Val Ser Thr Thr 
    610                 615                 620                 


Asp Lys Ile Ala Asp Ile Thr Ile Ile Ile Pro Tyr Ile Gly Pro Ala 
625                 630                 635                 640 


Leu Asn Ile Gly Asn Met Leu Tyr Lys Asp Asp Phe Val Gly Ala Leu 
                645                 650                 655     


Ile Phe Ser Gly Ala Val Ile Leu Leu Glu Phe Ile Pro Glu Ile Ala 
            660                 665                 670         


Ile Pro Val Leu Gly Thr Phe Ala Leu Val Ser Tyr Ile Ala Asn Lys 
        675                 680                 685             


Val Leu Thr Val Gln Thr Ile Asp Asn Ala Leu Ser Lys Arg Asn Glu 
    690                 695                 700                 


Lys Trp Asp Glu Val Tyr Lys Tyr Ile Val Thr Asn Trp Leu Ala Lys 
705                 710                 715                 720 


Val Asn Thr Gln Ile Asp Leu Ile Arg Lys Lys Met Lys Glu Ala Leu 
                725                 730                 735     


Glu Asn Gln Ala Glu Ala Thr Lys Ala Ile Ile Asn Tyr Gln Tyr Asn 
            740                 745                 750         


Gln Tyr Thr Glu Glu Glu Lys Asn Asn Ile Asn Phe Asn Ile Asp Asp 
        755                 760                 765             


Leu Ser Ser Lys Leu Asn Glu Ser Ile Asn Lys Ala Met Ile Asn Ile 
    770                 775                 780                 


Asn Lys Phe Leu Asn Gln Cys Ser Val Ser Tyr Leu Met Asn Ser Met 
785                 790                 795                 800 


Ile Pro Tyr Gly Val Lys Arg Leu Glu Asp Phe Asp Ala Ser Leu Lys 
                805                 810                 815     


Asp Ala Leu Leu Lys Tyr Ile Tyr Asp Asn Arg Gly Thr Leu Ile Gly 
            820                 825                 830         


Gln Val Asp Arg Leu Lys Asp Lys Val Asn Asn Thr Leu Ser Thr Asp 
        835                 840                 845             


Ile Pro Phe Gln Leu Ser Lys Tyr Val Asp Asn Gln Arg Leu Leu Ser 
    850                 855                 860                 


Thr Phe Thr Glu Tyr Ile Lys Asn Ile Ile Asn Thr Ser Ile Leu Asn 
865                 870                 875                 880 


Leu Arg Tyr Glu Ser Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser 
                885                 890                 895     


Lys Ile Asn Ile Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn 
            900                 905                 910         


Gln Ile Gln Leu Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu 
        915                 920                 925             


Lys Lys Ala Ile Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser 
    930                 935                 940                 


Phe Trp Ile Arg Ile Pro Lys Tyr Phe Lys Lys Ile Ser Leu Asn Asn 
945                 950                 955                 960 


Glu Tyr Thr Ile Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val 
                965                 970                 975     


Ser Leu Asn Tyr Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Lys Glu 
            980                 985                 990         


Ile Lys Gln Arg Val Val Phe Lys  Tyr Ser Gln Met Ile  Asn Ile Ser 
        995                 1000                 1005             


Asp Tyr  Ile Asn Arg Trp Ile  Phe Val Thr Ile Thr  Asn Asn Arg 
    1010                 1015                 1020             


Leu Asn  Lys Ser Lys Ile Tyr  Ile Asn Gly Arg Leu  Ile Asp Gln 
    1025                 1030                 1035             


Lys Pro  Ile Ser Asn Leu Gly  Asn Ile His Ala Ser  Asn Lys Ile 
    1040                 1045                 1050             


Met Phe  Lys Leu Asp Gly Cys  Arg Asp Thr His Arg  Tyr Ile Trp 
    1055                 1060                 1065             


Ile Lys  Tyr Phe Asn Leu Phe  Asp Lys Glu Leu Asn  Glu Lys Glu 
    1070                 1075                 1080             


Ile Lys  Asp Leu Tyr Asp Asn  Gln Ser Asn Ser Gly  Ile Leu Lys 
    1085                 1090                 1095             


Asp Phe  Trp Gly Asp Tyr Leu  Gln Tyr Asp Lys Pro  Tyr Tyr Met 
    1100                 1105                 1110             


Leu Asn  Leu Tyr Asp Pro Asn  Lys Tyr Val Asp Val  Asn Asn Val 
    1115                 1120                 1125             


Gly Ile  Arg Gly Tyr Met Tyr  Leu Lys Gly Pro Arg  Gly Ser Val 
    1130                 1135                 1140             


Met Thr  Thr Asn Ile Tyr Leu  Asn Ser Ser Leu Tyr  Arg Gly Thr 
    1145                 1150                 1155             


Lys Phe  Ile Ile Lys Lys Tyr  Ala Ser Gly Asn Lys  Asp Asn Ile 
    1160                 1165                 1170             


Val Arg  Asn Asn Asp Arg Val  Tyr Ile Asn Val Val  Val Lys Asn 
    1175                 1180                 1185             


Lys Glu  Tyr Arg Leu Ala Thr  Asn Ala Ser Gln Ala  Gly Val Glu 
    1190                 1195                 1200             


Lys Ile  Leu Ser Ala Leu Glu  Ile Pro Asp Val Gly  Asn Leu Ser 
    1205                 1210                 1215             


Gln Val  Val Val Met Lys Ser  Lys Asn Asp Lys Gly  Ile Thr Asn 
    1220                 1225                 1230             


Lys Cys  Lys Met Asn Leu Gln  Asp Asn Asn Gly Asn  Asp Ile Gly 
    1235                 1240                 1245             


Phe Ile  Gly Phe His Gln Phe  Asn Asn Ile Ala Lys  Leu Val Ala 
    1250                 1255                 1260             


Ser Asn  Trp Tyr Asn Arg Gln  Ile Glu Arg Ser Ser  Arg Thr Leu 
    1265                 1270                 1275             


Gly Cys  Ser Trp Glu Phe Ile  Pro Val Asp Asp Gly  Trp Gly Glu 
    1280                 1285                 1290             


Arg Pro  Leu 
    1295     


<210>  7
<211>  3888
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Engineered BoNT/A1

<400>  7
atgccattcg tcaacaagca attcaactac aaagacccag tcaacggcgt cgacatcgca       60

tacatcaaga ttccgaacgc cggtcaaatg cagccggtta aggcttttaa gatccacaac      120

aagatttggg ttatcccgga gcgtgacacc ttcacgaacc cggaagaagg cgatctgaac      180

ccgccaccgg aagcgaagca agtccctgtc agctactacg attcgacgta cctgagcacg      240

gataacgaaa aagataacta cctgaaaggt gtgaccaagc tgttcgaacg tatctacagc      300

acggatctgg gtcgcatgct gctgactagc attgttcgcg gtatcccgtt ctggggtggt      360

agcacgattg acaccgaact gaaggttatc gacactaact gcattaacgt tattcaaccg      420

gatggtagct atcgtagcga agagctgaat ctggtcatca ttggcccgag cgcagacatt      480

atccaattcg agtgcaagag ctttggtcac gaggttctga atctgacccg caatggctat      540

ggtagcaccc agtacattcg tttttcgccg gattttacct tcggctttga agagagcctg      600

gaggttgata ccaatccgtt gctgggtgcg ggcaaattcg ctaccgatcc ggctgtcacg      660

ctggcccatg aactgatcca cgcaggccac cgcctgtacg gcattgccat caacccaaac      720

cgtgtgttca aggttaatac gaatgcatac tacgagatga gcggcctgga agtcagcttc      780

gaagaactgc gcaccttcgg tggccatgac gctaaattca ttgacagctt gcaagagaat      840

gagttccgtc tgtactacta taacaaattc aaagacattg caagcacgtt gaacaaggcc      900

aaaagcatcg ttggtactac cgcgtcgttg cagtatatga agaatgtgtt taaagagaag      960

tacctgctgt ccgaggatac ctccggcaag tttagcgttg ataagctgaa gtttgacaaa     1020

ctgtacaaga tgctgaccga gatttacacc gaggacaact ttgtgaaatt cttcaaagtg     1080

ttgaatcgta aaacctatct gaattttgac aaagcggttt tcaagattaa catcgtgccg     1140

aaggtgaact acaccatcta tgacggtttt aacctgcgta acaccaacct ggcggcgaac     1200

tttaacggtc agaatacgga aatcaacaac atgaatttca cgaagttgaa gaacttcacg     1260

ggtctgttcg agttctataa gctgctgtgc gtgcgcggta tcatcaccag caaaaccaaa     1320

agcctggaca aaggctacaa caaggcgctg aatgacctgt gcattaaggt aaacaattgg     1380

gatctgttct tttcgccatc cgaagataat tttaccaacg acctgaacaa gggtgaagaa     1440

atcaccagcg atacgaatat tgaagcagcg gaagagaata tcagcctgga tctgatccag     1500

cagtactatc tgacctttaa cttcgacaat gaaccggaga acattagcat tgagaatctg     1560

agcagcgaca ttatcggtca gctggaactg atgccgaata tcgaacgttt cccgaacggc     1620

aaaaagtacg agctggacaa gtacactatg ttccattacc tgcgtgcaca ggagtttgaa     1680

cacggtaaaa gccgtatcgc gctgaccaac agcgttaacg aggccctgct gaacccgagc     1740

cgtgtctata ccttcttcag cagcgactat gttaagaaag tgaacaaagc cactgaggcc     1800

gcgatgttcc tgggctgggt ggaacagctg gtatatgact tcacggacga gacgagcgaa     1860

gtgagcacta ccgacaaaat tgctgatatt accatcatta tcccgtatat tggtccggca     1920

ctgaacattg gcaacatgct gtacaaagac gattttgtgg gtgccctgat cttctccggt     1980

gccgtgattc tgctggagtt cattccggag attgcgatcc cggtgttggg taccttcgcg     2040

ctggtgtcct acatcgcgaa taaggttctg acggttcaga ccatcgataa cgcgctgtcg     2100

aaacgtaatg aaaaatggga cgaggtttac aaatacattg ttacgaattg gctggcgaaa     2160

gtcaataccc agatcgacct gatccgtaag aaaatgaaag aggcgctgga gaatcaggcg     2220

gaggccacca aagcaattat caactaccaa tacaaccagt acacggaaga agagaagaat     2280

aacattaact tcaatatcga tgatttgagc agcaagctga atgaatctat caacaaagcg     2340

atgatcaata tcaacaagtt tttgaatcag tgtagcgttt cgtacctgat gaatagcatg     2400

attccgtatg gcgtcaaacg tctggaggac ttcgacgcca gcctgaaaga tgcgttgctg     2460

aaatacattt acgacaatcg tggtacgctg attggccaag ttgaccgctt gaaagacaaa     2520

gttaacaata ccctgagcac cgacatccca tttcaactga gcaagtatgt tgataatcaa     2580

cgtctgttga gcactttcac cgagtatatc aaaaacatca tcaatactag cattctgaac     2640

ctgcgttacg agagcaatca tctgattgat ctgagccgtt atgctagcaa gatcaacatc     2700

ggtagcaagg tcaattttga cccgatcgat aagaaccaga tccagctgtt taatctggaa     2760

tcgagcaaaa ttgaggttat cctgaaaaag gccattgtct acaactccat gtacgagaat     2820

ttctccacca gcttctggat tcgcatcccg aaatacttca acaagattag cctgaacaac     2880

gagtatacta tcatcaactg tatggagaac aacagcggtt ggaaggtgtc tctgaactat     2940

ggtgagatca tttggacctt gcaggacacc aaagagatca agcagcgcgt cgtgttcaag     3000

tactctcaaa tgatcaacat ttccgattac attaatcgtt ggatcttcgt gaccattacg     3060

aataaccgtc tgaagaagag caagatttac atcaatggtc gcttgatcga tcagaaaccg     3120

attagcaacc tgggtaatat ccacgcaagc aacaagatta tgttcaaatt ggacggttgc     3180

cgcgataccc atcgttatat ctggatcaag tatttcaacc tgtttgataa agaactgaat     3240

gagaaggaga tcaaagattt gtatgacaac caatctaaca gcggcatttt gaaggacttc     3300

tggggcgatt atctgcaata cgataagccg tactatatgc tgaacctgta tgatccgaac     3360

aaatatgtgg atgtcaataa tgtgggtatt cgtggttaca tgtatttgaa gggtccgcgt     3420

ggcagcgtta tgacgaccaa catttacctg aactctagcc tgtaccgtgg tacgaaattc     3480

atcattaaga aatatgccag cggcaacaaa gataacattg tgcgtaataa cgatcgtgtc     3540

tacatcaacg tggtcgtgaa gaataaagag taccgtctgg cgaccaacgc ttcgcaggcg     3600

ggtgttgaga aaattctgag cgcgttggag atccctgatg tcggtaatct gagccaagtc     3660

gtggttatga agagcaagaa cgacaagggt atcactaaca agtgcaagat gaacctgcaa     3720

gacaacaatg gtaacgacat cggctttatt ggtttccacc agttcaacaa tattgctaaa     3780

ctggtagcga gcaattggta caatcgtcag attgagcgca gcagccgtac tttgggctgt     3840

agctgggagt ttatcccggt cgatgatggt tggggcgaac gtccgctg                  3888


<210>  8
<211>  1296
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Engineered BoNT/A1

<400>  8

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             


Ala Leu Asn Asp Leu Cys Ile Lys Val Asn Asn Trp Asp Leu Phe Phe 
    450                 455                 460                 


Ser Pro Ser Glu Asp Asn Phe Thr Asn Asp Leu Asn Lys Gly Glu Glu 
465                 470                 475                 480 


Ile Thr Ser Asp Thr Asn Ile Glu Ala Ala Glu Glu Asn Ile Ser Leu 
                485                 490                 495     


Asp Leu Ile Gln Gln Tyr Tyr Leu Thr Phe Asn Phe Asp Asn Glu Pro 
            500                 505                 510         


Glu Asn Ile Ser Ile Glu Asn Leu Ser Ser Asp Ile Ile Gly Gln Leu 
        515                 520                 525             


Glu Leu Met Pro Asn Ile Glu Arg Phe Pro Asn Gly Lys Lys Tyr Glu 
    530                 535                 540                 


Leu Asp Lys Tyr Thr Met Phe His Tyr Leu Arg Ala Gln Glu Phe Glu 
545                 550                 555                 560 


His Gly Lys Ser Arg Ile Ala Leu Thr Asn Ser Val Asn Glu Ala Leu 
                565                 570                 575     


Leu Asn Pro Ser Arg Val Tyr Thr Phe Phe Ser Ser Asp Tyr Val Lys 
            580                 585                 590         


Lys Val Asn Lys Ala Thr Glu Ala Ala Met Phe Leu Gly Trp Val Glu 
        595                 600                 605             


Gln Leu Val Tyr Asp Phe Thr Asp Glu Thr Ser Glu Val Ser Thr Thr 
    610                 615                 620                 


Asp Lys Ile Ala Asp Ile Thr Ile Ile Ile Pro Tyr Ile Gly Pro Ala 
625                 630                 635                 640 


Leu Asn Ile Gly Asn Met Leu Tyr Lys Asp Asp Phe Val Gly Ala Leu 
                645                 650                 655     


Ile Phe Ser Gly Ala Val Ile Leu Leu Glu Phe Ile Pro Glu Ile Ala 
            660                 665                 670         


Ile Pro Val Leu Gly Thr Phe Ala Leu Val Ser Tyr Ile Ala Asn Lys 
        675                 680                 685             


Val Leu Thr Val Gln Thr Ile Asp Asn Ala Leu Ser Lys Arg Asn Glu 
    690                 695                 700                 


Lys Trp Asp Glu Val Tyr Lys Tyr Ile Val Thr Asn Trp Leu Ala Lys 
705                 710                 715                 720 


Val Asn Thr Gln Ile Asp Leu Ile Arg Lys Lys Met Lys Glu Ala Leu 
                725                 730                 735     


Glu Asn Gln Ala Glu Ala Thr Lys Ala Ile Ile Asn Tyr Gln Tyr Asn 
            740                 745                 750         


Gln Tyr Thr Glu Glu Glu Lys Asn Asn Ile Asn Phe Asn Ile Asp Asp 
        755                 760                 765             


Leu Ser Ser Lys Leu Asn Glu Ser Ile Asn Lys Ala Met Ile Asn Ile 
    770                 775                 780                 


Asn Lys Phe Leu Asn Gln Cys Ser Val Ser Tyr Leu Met Asn Ser Met 
785                 790                 795                 800 


Ile Pro Tyr Gly Val Lys Arg Leu Glu Asp Phe Asp Ala Ser Leu Lys 
                805                 810                 815     


Asp Ala Leu Leu Lys Tyr Ile Tyr Asp Asn Arg Gly Thr Leu Ile Gly 
            820                 825                 830         


Gln Val Asp Arg Leu Lys Asp Lys Val Asn Asn Thr Leu Ser Thr Asp 
        835                 840                 845             


Ile Pro Phe Gln Leu Ser Lys Tyr Val Asp Asn Gln Arg Leu Leu Ser 
    850                 855                 860                 


Thr Phe Thr Glu Tyr Ile Lys Asn Ile Ile Asn Thr Ser Ile Leu Asn 
865                 870                 875                 880 


Leu Arg Tyr Glu Ser Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser 
                885                 890                 895     


Lys Ile Asn Ile Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn 
            900                 905                 910         


Gln Ile Gln Leu Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu 
        915                 920                 925             


Lys Lys Ala Ile Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser 
    930                 935                 940                 


Phe Trp Ile Arg Ile Pro Lys Tyr Phe Asn Lys Ile Ser Leu Asn Asn 
945                 950                 955                 960 


Glu Tyr Thr Ile Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val 
                965                 970                 975     


Ser Leu Asn Tyr Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Lys Glu 
            980                 985                 990         


Ile Lys Gln Arg Val Val Phe Lys  Tyr Ser Gln Met Ile  Asn Ile Ser 
        995                 1000                 1005             


Asp Tyr  Ile Asn Arg Trp Ile  Phe Val Thr Ile Thr  Asn Asn Arg 
    1010                 1015                 1020             


Leu Lys  Lys Ser Lys Ile Tyr  Ile Asn Gly Arg Leu  Ile Asp Gln 
    1025                 1030                 1035             


Lys Pro  Ile Ser Asn Leu Gly  Asn Ile His Ala Ser  Asn Lys Ile 
    1040                 1045                 1050             


Met Phe  Lys Leu Asp Gly Cys  Arg Asp Thr His Arg  Tyr Ile Trp 
    1055                 1060                 1065             


Ile Lys  Tyr Phe Asn Leu Phe  Asp Lys Glu Leu Asn  Glu Lys Glu 
    1070                 1075                 1080             


Ile Lys  Asp Leu Tyr Asp Asn  Gln Ser Asn Ser Gly  Ile Leu Lys 
    1085                 1090                 1095             


Asp Phe  Trp Gly Asp Tyr Leu  Gln Tyr Asp Lys Pro  Tyr Tyr Met 
    1100                 1105                 1110             


Leu Asn  Leu Tyr Asp Pro Asn  Lys Tyr Val Asp Val  Asn Asn Val 
    1115                 1120                 1125             


Gly Ile  Arg Gly Tyr Met Tyr  Leu Lys Gly Pro Arg  Gly Ser Val 
    1130                 1135                 1140             


Met Thr  Thr Asn Ile Tyr Leu  Asn Ser Ser Leu Tyr  Arg Gly Thr 
    1145                 1150                 1155             


Lys Phe  Ile Ile Lys Lys Tyr  Ala Ser Gly Asn Lys  Asp Asn Ile 
    1160                 1165                 1170             


Val Arg  Asn Asn Asp Arg Val  Tyr Ile Asn Val Val  Val Lys Asn 
    1175                 1180                 1185             


Lys Glu  Tyr Arg Leu Ala Thr  Asn Ala Ser Gln Ala  Gly Val Glu 
    1190                 1195                 1200             


Lys Ile  Leu Ser Ala Leu Glu  Ile Pro Asp Val Gly  Asn Leu Ser 
    1205                 1210                 1215             


Gln Val  Val Val Met Lys Ser  Lys Asn Asp Lys Gly  Ile Thr Asn 
    1220                 1225                 1230             


Lys Cys  Lys Met Asn Leu Gln  Asp Asn Asn Gly Asn  Asp Ile Gly 
    1235                 1240                 1245             


Phe Ile  Gly Phe His Gln Phe  Asn Asn Ile Ala Lys  Leu Val Ala 
    1250                 1255                 1260             


Ser Asn  Trp Tyr Asn Arg Gln  Ile Glu Arg Ser Ser  Arg Thr Leu 
    1265                 1270                 1275             


Gly Cys  Ser Trp Glu Phe Ile  Pro Val Asp Asp Gly  Trp Gly Glu 
    1280                 1285                 1290             


Arg Pro  Leu 
    1295     


<210>  9
<211>  3888
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Engineered BoNT/A1

<400>  9
atgccattcg tcaacaagca attcaactac aaagacccag tcaacggcgt cgacatcgca       60

tacatcaaga ttccgaacgc cggtcaaatg cagccggtta aggcttttaa gatccacaac      120

aagatttggg ttatcccgga gcgtgacacc ttcacgaacc cggaagaagg cgatctgaac      180

ccgccaccgg aagcgaagca agtccctgtc agctactacg attcgacgta cctgagcacg      240

gataacgaaa aagataacta cctgaaaggt gtgaccaagc tgttcgaacg tatctacagc      300

acggatctgg gtcgcatgct gctgactagc attgttcgcg gtatcccgtt ctggggtggt      360

agcacgattg acaccgaact gaaggttatc gacactaact gcattaacgt tattcaaccg      420

gatggtagct atcgtagcga agagctgaat ctggtcatca ttggcccgag cgcagacatt      480

atccaattcg agtgcaagag ctttggtcac gaggttctga atctgacccg caatggctat      540

ggtagcaccc agtacattcg tttttcgccg gattttacct tcggctttga agagagcctg      600

gaggttgata ccaatccgtt gctgggtgcg ggcaaattcg ctaccgatcc ggctgtcacg      660

ctggcccatg aactgatcca cgcaggccac cgcctgtacg gcattgccat caacccaaac      720

cgtgtgttca aggttaatac gaatgcatac tacgagatga gcggcctgga agtcagcttc      780

gaagaactgc gcaccttcgg tggccatgac gctaaattca ttgacagctt gcaagagaat      840

gagttccgtc tgtactacta taacaaattc aaagacattg caagcacgtt gaacaaggcc      900

aaaagcatcg ttggtactac cgcgtcgttg cagtatatga agaatgtgtt taaagagaag      960

tacctgctgt ccgaggatac ctccggcaag tttagcgttg ataagctgaa gtttgacaaa     1020

ctgtacaaga tgctgaccga gatttacacc gaggacaact ttgtgaaatt cttcaaagtg     1080

ttgaatcgta aaacctatct gaattttgac aaagcggttt tcaagattaa catcgtgccg     1140

aaggtgaact acaccatcta tgacggtttt aacctgcgta acaccaacct ggcggcgaac     1200

tttaacggtc agaatacgga aatcaacaac atgaatttca cgaagttgaa gaacttcacg     1260

ggtctgttcg agttctataa gctgctgtgc gtgcgcggta tcatcaccag caaaaccaaa     1320

agcctggaca aaggctacaa caaggcgctg aatgacctgt gcattaaggt aaacaattgg     1380

gatctgttct tttcgccatc cgaagataat tttaccaacg acctgaacaa gggtgaagaa     1440

atcaccagcg atacgaatat tgaagcagcg gaagagaata tcagcctgga tctgatccag     1500

cagtactatc tgacctttaa cttcgacaat gaaccggaga acattagcat tgagaatctg     1560

agcagcgaca ttatcggtca gctggaactg atgccgaata tcgaacgttt cccgaacggc     1620

aaaaagtacg agctggacaa gtacactatg ttccattacc tgcgtgcaca ggagtttgaa     1680

cacggtaaaa gccgtatcgc gctgaccaac agcgttaacg aggccctgct gaacccgagc     1740

cgtgtctata ccttcttcag cagcgactat gttaagaaag tgaacaaagc cactgaggcc     1800

gcgatgttcc tgggctgggt ggaacagctg gtatatgact tcacggacga gacgagcgaa     1860

gtgagcacta ccgacaaaat tgctgatatt accatcatta tcccgtatat tggtccggca     1920

ctgaacattg gcaacatgct gtacaaagac gattttgtgg gtgccctgat cttctccggt     1980

gccgtgattc tgctggagtt cattccggag attgcgatcc cggtgttggg taccttcgcg     2040

ctggtgtcct acatcgcgaa taaggttctg acggttcaga ccatcgataa cgcgctgtcg     2100

aaacgtaatg aaaaatggga cgaggtttac aaatacattg ttacgaattg gctggcgaaa     2160

gtcaataccc agatcgacct gatccgtaag aaaatgaaag aggcgctgga gaatcaggcg     2220

gaggccacca aagcaattat caactaccaa tacaaccagt acacggaaga agagaagaat     2280

aacattaact tcaatatcga tgatttgagc agcaagctga atgaatctat caacaaagcg     2340

atgatcaata tcaacaagtt tttgaatcag tgtagcgttt cgtacctgat gaatagcatg     2400

attccgtatg gcgtcaaacg tctggaggac ttcgacgcca gcctgaaaga tgcgttgctg     2460

aaatacattt acgacaatcg tggtacgctg attggccaag ttgaccgctt gaaagacaaa     2520

gttaacaata ccctgagcac cgacatccca tttcaactga gcaagtatgt tgataatcaa     2580

cgtctgttga gcactttcac cgagtatatc aaaaacatca tcaatactag cattctgaac     2640

ctgcgttacg agagcaatca tctgattgat ctgagccgtt atgcaagcaa gatcaacatc     2700

ggtagcaagg tcaattttga cccgatcgat aagaaccaga tccagctgtt taatctggaa     2760

tcgagcaaaa ttgaggttat cctgaaaaac gccattgtct acaactccat gtacgagaat     2820

ttctccacca gcttctggat tcgcatcccg aaatacttca acagcattag cctgaacaac     2880

gagtatacta tcatcaactg tatggagaac aacagcggtt ggaaggtgtc tctgaactat     2940

ggtgagatca tttggacctt gcaggacacc caagagatca agcagcgcgt cgtgttcaag     3000

tactctcaaa tgatcaacat ttccgattac attaatcgtt ggatcttcgt gaccattacg     3060

aataaccgtc tgaataacag caagatttac atcaatggtc gcttgatcga tcagaaaccg     3120

attagcaacc tgggtaatat ccacgcaagc aacaacatta tgttcaaatt ggacggttgc     3180

cgcgataccc atcgttatat ctggatcaag tatttcaacc tgtttgataa agaactgaat     3240

gagaaggaga tcaaagattt gtatgacaac caatctaaca gcggcatttt gaaggacttc     3300

tggggcgatt atctgcaata cgataagccg tactatatgc tgaacctgta tgatccgaac     3360

aaatatgtgg atgtcaataa tgtgggtatt cgtggttaca tgtatttgaa gggtccgcgt     3420

ggcagcgtta tgacgaccaa catttacctg aactctagcc tgtaccgtgg tacgaaattc     3480

atcattaaga aatatgccag cggcaacaaa gataacattg tgcgtaataa cgatcgtgtc     3540

tacatcaacg tggtcgtgaa gcgtaaagag taccgtctgg cgaccaacgc ttcgcaggcg     3600

ggtgttgaga aaattctgag cgcgttggag atccctcgtg tccgtcgtct gagccaagtc     3660

gtggttatga agagcaagaa cgaccagggt atcactaaca agtgcaagat gaacctgcaa     3720

gaccgtcgtg gtaacgacat cggctttatt ggtttccacc agttcaacaa tattgctaaa     3780

ctggtagcga gcaattggta caatcgtcag attgagcgcc gtagccgtcg tttgggctgt     3840

agctgggagt ttatcccggt cgatgatggt tggggcgaac gtccgctg                  3888


<210>  10
<211>  1296
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Engineered BoNT/A1

<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 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             


Ala Leu Asn Asp Leu Cys Ile Lys Val Asn Asn Trp Asp Leu Phe Phe 
    450                 455                 460                 


Ser Pro Ser Glu Asp Asn Phe Thr Asn Asp Leu Asn Lys Gly Glu Glu 
465                 470                 475                 480 


Ile Thr Ser Asp Thr Asn Ile Glu Ala Ala Glu Glu Asn Ile Ser Leu 
                485                 490                 495     


Asp Leu Ile Gln Gln Tyr Tyr Leu Thr Phe Asn Phe Asp Asn Glu Pro 
            500                 505                 510         


Glu Asn Ile Ser Ile Glu Asn Leu Ser Ser Asp Ile Ile Gly Gln Leu 
        515                 520                 525             


Glu Leu Met Pro Asn Ile Glu Arg Phe Pro Asn Gly Lys Lys Tyr Glu 
    530                 535                 540                 


Leu Asp Lys Tyr Thr Met Phe His Tyr Leu Arg Ala Gln Glu Phe Glu 
545                 550                 555                 560 


His Gly Lys Ser Arg Ile Ala Leu Thr Asn Ser Val Asn Glu Ala Leu 
                565                 570                 575     


Leu Asn Pro Ser Arg Val Tyr Thr Phe Phe Ser Ser Asp Tyr Val Lys 
            580                 585                 590         


Lys Val Asn Lys Ala Thr Glu Ala Ala Met Phe Leu Gly Trp Val Glu 
        595                 600                 605             


Gln Leu Val Tyr Asp Phe Thr Asp Glu Thr Ser Glu Val Ser Thr Thr 
    610                 615                 620                 


Asp Lys Ile Ala Asp Ile Thr Ile Ile Ile Pro Tyr Ile Gly Pro Ala 
625                 630                 635                 640 


Leu Asn Ile Gly Asn Met Leu Tyr Lys Asp Asp Phe Val Gly Ala Leu 
                645                 650                 655     


Ile Phe Ser Gly Ala Val Ile Leu Leu Glu Phe Ile Pro Glu Ile Ala 
            660                 665                 670         


Ile Pro Val Leu Gly Thr Phe Ala Leu Val Ser Tyr Ile Ala Asn Lys 
        675                 680                 685             


Val Leu Thr Val Gln Thr Ile Asp Asn Ala Leu Ser Lys Arg Asn Glu 
    690                 695                 700                 


Lys Trp Asp Glu Val Tyr Lys Tyr Ile Val Thr Asn Trp Leu Ala Lys 
705                 710                 715                 720 


Val Asn Thr Gln Ile Asp Leu Ile Arg Lys Lys Met Lys Glu Ala Leu 
                725                 730                 735     


Glu Asn Gln Ala Glu Ala Thr Lys Ala Ile Ile Asn Tyr Gln Tyr Asn 
            740                 745                 750         


Gln Tyr Thr Glu Glu Glu Lys Asn Asn Ile Asn Phe Asn Ile Asp Asp 
        755                 760                 765             


Leu Ser Ser Lys Leu Asn Glu Ser Ile Asn Lys Ala Met Ile Asn Ile 
    770                 775                 780                 


Asn Lys Phe Leu Asn Gln Cys Ser Val Ser Tyr Leu Met Asn Ser Met 
785                 790                 795                 800 


Ile Pro Tyr Gly Val Lys Arg Leu Glu Asp Phe Asp Ala Ser Leu Lys 
                805                 810                 815     


Asp Ala Leu Leu Lys Tyr Ile Tyr Asp Asn Arg Gly Thr Leu Ile Gly 
            820                 825                 830         


Gln Val Asp Arg Leu Lys Asp Lys Val Asn Asn Thr Leu Ser Thr Asp 
        835                 840                 845             


Ile Pro Phe Gln Leu Ser Lys Tyr Val Asp Asn Gln Arg Leu Leu Ser 
    850                 855                 860                 


Thr Phe Thr Glu Tyr Ile Lys Asn Ile Ile Asn Thr Ser Ile Leu Asn 
865                 870                 875                 880 


Leu Arg Tyr Glu Ser Asn His Leu Ile Asp Leu Ser Arg Tyr Ala Ser 
                885                 890                 895     


Lys Ile Asn Ile Gly Ser Lys Val Asn Phe Asp Pro Ile Asp Lys Asn 
            900                 905                 910         


Gln Ile Gln Leu Phe Asn Leu Glu Ser Ser Lys Ile Glu Val Ile Leu 
        915                 920                 925             


Lys Asn Ala Ile Val Tyr Asn Ser Met Tyr Glu Asn Phe Ser Thr Ser 
    930                 935                 940                 


Phe Trp Ile Arg Ile Pro Lys Tyr Phe Asn Ser Ile Ser Leu Asn Asn 
945                 950                 955                 960 


Glu Tyr Thr Ile Ile Asn Cys Met Glu Asn Asn Ser Gly Trp Lys Val 
                965                 970                 975     


Ser Leu Asn Tyr Gly Glu Ile Ile Trp Thr Leu Gln Asp Thr Gln Glu 
            980                 985                 990         


Ile Lys Gln Arg Val Val Phe Lys  Tyr Ser Gln Met Ile  Asn Ile Ser 
        995                 1000                 1005             


Asp Tyr  Ile Asn Arg Trp Ile  Phe Val Thr Ile Thr  Asn Asn Arg 
    1010                 1015                 1020             


Leu Asn  Asn Ser Lys Ile Tyr  Ile Asn Gly Arg Leu  Ile Asp Gln 
    1025                 1030                 1035             


Lys Pro  Ile Ser Asn Leu Gly  Asn Ile His Ala Ser  Asn Asn Ile 
    1040                 1045                 1050             


Met Phe  Lys Leu Asp Gly Cys  Arg Asp Thr His Arg  Tyr Ile Trp 
    1055                 1060                 1065             


Ile Lys  Tyr Phe Asn Leu Phe  Asp Lys Glu Leu Asn  Glu Lys Glu 
    1070                 1075                 1080             


Ile Lys  Asp Leu Tyr Asp Asn  Gln Ser Asn Ser Gly  Ile Leu Lys 
    1085                 1090                 1095             


Asp Phe  Trp Gly Asp Tyr Leu  Gln Tyr Asp Lys Pro  Tyr Tyr Met 
    1100                 1105                 1110             


Leu Asn  Leu Tyr Asp Pro Asn  Lys Tyr Val Asp Val  Asn Asn Val 
    1115                 1120                 1125             


Gly Ile  Arg Gly Tyr Met Tyr  Leu Lys Gly Pro Arg  Gly Ser Val 
    1130                 1135                 1140             


Met Thr  Thr Asn Ile Tyr Leu  Asn Ser Ser Leu Tyr  Arg Gly Thr 
    1145                 1150                 1155             


Lys Phe  Ile Ile Lys Lys Tyr  Ala Ser Gly Asn Lys  Asp Asn Ile 
    1160                 1165                 1170             


Val Arg  Asn Asn Asp Arg Val  Tyr Ile Asn Val Val  Val Lys Arg 
    1175                 1180                 1185             


Lys Glu  Tyr Arg Leu Ala Thr  Asn Ala Ser Gln Ala  Gly Val Glu 
    1190                 1195                 1200             


Lys Ile  Leu Ser Ala Leu Glu  Ile Pro Arg Val Arg  Arg Leu Ser 
    1205                 1210                 1215             


Gln Val  Val Val Met Lys Ser  Lys Asn Asp Gln Gly  Ile Thr Asn 
    1220                 1225                 1230             


Lys Cys  Lys Met Asn Leu Gln  Asp Arg Arg Gly Asn  Asp Ile Gly 
    1235                 1240                 1245             


Phe Ile  Gly Phe His Gln Phe  Asn Asn Ile Ala Lys  Leu Val Ala 
    1250                 1255                 1260             


Ser Asn  Trp Tyr Asn Arg Gln  Ile Glu Arg Arg Ser  Arg Arg Leu 
    1265                 1270                 1275             


Gly Cys  Ser Trp Glu Phe Ile  Pro Val Asp Asp Gly  Trp Gly Glu 
    1280                 1285                 1290             


Arg Pro  Leu 
    1295     


