                         SEQUENCE LISTING

<110>  University of Rochester
       Zeng, Mingtao
       Pichichero, Michael E.
       Xu, Qingfu
 
<120>  VACCINE AGAINST BOTULISM

<130>  176/62351

<150>  60/954,921
<151>  2007-08-09

<160>  8     

<170>  PatentIn version 3.5

<210>  1
<211>  1299
<212>  DNA
<213>  Artificial

<220>
<223>  wildtype sequence encoding 50kD heavy chain fragment of 
       Clostridium botulinum neurotoxin C

<400>  1
gatataatta atgaatattt caataatatt aatgattcaa aaattttgag cctacaaaac       60

agaaaaaata ctttagtgga tacatcagga tataatgcag aagtgagtga agaaggcgat      120

gttcagctta atccaatatt tccatttgac tttaaattag gtagttcagg ggaggataga      180

ggtaaagtta tagtaaccca gaatgaaaat attgtatata attctatgta tgaaagtttt      240

agcattagtt tttggattag aataaataaa tgggtaagta atttacctgg atatactata      300

attgatagtg ttaaaaataa ctcaggttgg agtataggta ttattagtaa ttttttagta      360

tttactttaa aacaaaatga agatagtgaa caaagtataa attttagtta tgatatatca      420

aataatgctc ctggatacaa taaatggttt tttgtaactg ttactaacaa tatgatggga      480

aatatgaaga tttatataaa tggaaaatta atagatacta taaaagttaa agaactaact      540

ggaattaatt ttagcaaaac tataacattt gaaataaata aaattccaga taccggtttg      600

attacttcag attctgataa catcaatatg tggataagag atttttatat atttgctaaa      660

gaattagatg gtaaagatat taatatatta tttaatagct tgcaatatac taatgttgta      720

aaagattatt ggggaaatga tttaagatat aataaagaat attatatggt taatatagat      780

tatttaaata gatatatgta tgcgaactca cgacaaattg tttttaatac acgtagaaat      840

aataatgact tcaatgaagg atataaaatt ataataaaaa gaatcagagg aaatacaaat      900

gatactagag tacgaggagg agatatttta tattttgata tgacaattaa taacaaagca      960

tataatttgt ttatgaagaa tgaaactatg tatgcagata atcatagtac tgaagatata     1020

tatgctatag gtttaagaga acaaacaaag gatataaatg ataatattat atttcaaata     1080

caaccaatga ataatactta ttattacgca tctcaaatat ttaaatcaaa ttttaatgga     1140

gaaaatattt ctggaatatg ttcaataggt acttatcgtt ttagacttgg aggtgattgg     1200

tatagacaca attatttggt gcctactgtg aagcaaggaa attatgcttc attattagaa     1260

tcaacatcaa ctcattgggg ttttgtacct gtaagtgaa                            1299


<210>  2
<211>  1302
<212>  DNA
<213>  Artificial

<220>
<223>  codon-enhanced sequence encoding 50kD heavy chain fragment of 
       Clostridium botulinum neurotoxin C

<400>  2
gacattatca acgagtactt caataacatc aatgacagca agatcctgtc cctgcaaaat       60

cggaaaaaca ctctggtgga caccagcgga tataacgctg aggtgagcga agaaggcgat      120

gtgcaactga acccaatctt cccctttgat tttaagctgg gctcctccgg cgaggatagg      180

gggaaagtca tcgtcaccca gaatgaaaac atcgtctaca atagcatgta cgagagcttc      240

agcatctcct tctggatcag aattaacaaa tgggtcagca acctgccagg atataccatc      300

atcgacagcg tgaagaacaa ctccgggtgg tccatcggga ttatctccaa ttttctggtg      360

ttcactctga aacaaaacga agatagcgaa cagagcatca atttctccta cgacatttcc      420

aacaatgcac cagggtataa caagtggttc tttgtcactg tcaccaacaa catgatgggg      480

aacatgaaga tttacatcaa cggaaaactg attgatacta ttaaggtcaa ggaactcacc      540

ggcattaact tctccaagac aattacattt gagatcaata agattccaga caccggactc      600

attactagcg actccgacaa tatcaatatg tggattaggg acttctacat ctttgctaaa      660

gaactggatg gcaaggatat taacattctc ttcaactccc tccaatacac aaacgtcgtc      720

aaagactatt ggggcaacga cctgagatac aacaaagagt attacatggt caacatcgat      780

tacctgaaca gatatatgta cgccaacagc aggcaaattg tgttcaacac acggaggaat      840

aacaatgatt tcaacgaagg ctataagatc atcatcaaaa gaatcagggg aaacactaat      900

gacactaggg tcagaggcgg cgacattctg tattttgaca tgactatcaa caataaggcc      960

tacaacctgt ttatgaaaaa cgagacaatg tatgctgata accacagcac agaagatatt     1020

tacgcaatcg gcctgaggga gcaaaccaaa gacattaacg ataatatcat tttccagatc     1080

cagccaatga ataataccta ctactacgca agccaaattt tcaagagcaa ctttaacgga     1140

gagaacatca gcggaatctg cagcattggg acctacaggt ttagactcgg gggagactgg     1200

tatagacata attacctcgt gcctaccgtc aagcagggaa attatgccag cctcctcgaa     1260

agcacttcca cccattgggg atttgtcccc gtctccgagt ga                        1302


<210>  3
<211>  433
<212>  PRT
<213>  Artificial

<220>
<223>  50kD heavy chain fragment of Clostridium botulinum neurotoxin C

<400>  3

Asp Ile Ile Asn Glu Tyr Phe Asn Asn Ile Asn Asp Ser Lys Ile Leu 
1               5                   10                  15      


Ser Leu Gln Asn Arg Lys Asn Thr Leu Val Asp Thr Ser Gly Tyr Asn 
            20                  25                  30          


Ala Glu Val Ser Glu Glu Gly Asp Val Gln Leu Asn Pro Ile Phe Pro 
        35                  40                  45              


Phe Asp Phe Lys Leu Gly Ser Ser Gly Glu Asp Arg Gly Lys Val Ile 
    50                  55                  60                  


Val Thr Gln Asn Glu Asn Ile Val Tyr Asn Ser Met Tyr Glu Ser Phe 
65                  70                  75                  80  


Ser Ile Ser Phe Trp Ile Arg Ile Asn Lys Trp Val Ser Asn Leu Pro 
                85                  90                  95      


Gly Tyr Thr Ile Ile Asp Ser Val Lys Asn Asn Ser Gly Trp Ser Ile 
            100                 105                 110         


Gly Ile Ile Ser Asn Phe Leu Val Phe Thr Leu Lys Gln Asn Glu Asp 
        115                 120                 125             


Ser Glu Gln Ser Ile Asn Phe Ser Tyr Asp Ile Ser Asn Asn Ala Pro 
    130                 135                 140                 


Gly Tyr Asn Lys Trp Phe Phe Val Thr Val Thr Asn Asn Met Met Gly 
145                 150                 155                 160 


Asn Met Lys Ile Tyr Ile Asn Gly Lys Leu Ile Asp Thr Ile Lys Val 
                165                 170                 175     


Lys Glu Leu Thr Gly Ile Asn Phe Ser Lys Thr Ile Thr Phe Glu Ile 
            180                 185                 190         


Asn Lys Ile Pro Asp Thr Gly Leu Ile Thr Ser Asp Ser Asp Asn Ile 
        195                 200                 205             


Asn Met Trp Ile Arg Asp Phe Tyr Ile Phe Ala Lys Glu Leu Asp Gly 
    210                 215                 220                 


Lys Asp Ile Asn Ile Leu Phe Asn Ser Leu Gln Tyr Thr Asn Val Val 
225                 230                 235                 240 


Lys Asp Tyr Trp Gly Asn Asp Leu Arg Tyr Asn Lys Glu Tyr Tyr Met 
                245                 250                 255     


Val Asn Ile Asp Tyr Leu Asn Arg Tyr Met Tyr Ala Asn Ser Arg Gln 
            260                 265                 270         


Ile Val Phe Asn Thr Arg Arg Asn Asn Asn Asp Phe Asn Glu Gly Tyr 
        275                 280                 285             


Lys Ile Ile Ile Lys Arg Ile Arg Gly Asn Thr Asn Asp Thr Arg Val 
    290                 295                 300                 


Arg Gly Gly Asp Ile Leu Tyr Phe Asp Met Thr Ile Asn Asn Lys Ala 
305                 310                 315                 320 


Tyr Asn Leu Phe Met Lys Asn Glu Thr Met Tyr Ala Asp Asn His Ser 
                325                 330                 335     


Thr Glu Asp Ile Tyr Ala Ile Gly Leu Arg Glu Gln Thr Lys Asp Ile 
            340                 345                 350         


Asn Asp Asn Ile Ile Phe Gln Ile Gln Pro Met Asn Asn Thr Tyr Tyr 
        355                 360                 365             


Tyr Ala Ser Gln Ile Phe Lys Ser Asn Phe Asn Gly Glu Asn Ile Ser 
    370                 375                 380                 


Gly Ile Cys Ser Ile Gly Thr Tyr Arg Phe Arg Leu Gly Gly Asp Trp 
385                 390                 395                 400 


Tyr Arg His Asn Tyr Leu Val Pro Thr Val Lys Gln Gly Asn Tyr Ala 
                405                 410                 415     


Ser Leu Leu Glu Ser Thr Ser Thr His Trp Gly Phe Val Pro Val Ser 
            420                 425                 430         


Glu 
    


<210>  4
<211>  1311
<212>  DNA
<213>  Artificial

<220>
<223>  wildtype sequence encoding 50kD heavy chain fragment of 
       Clostridium botulinum neurotoxin A

<400>  4
agattattat ctacatttac tgaatatatt aagaatatta ttaatacttc tatattgaat       60

ttaagatatg aaagtaatca tttaatagac ttatctaggt atgcatcaaa aataaatatt      120

ggtagtaaag taaattttga tccaatagat aaaaatcaaa ttcaattatt taatttagaa      180

agtagtaaaa ttgaggtaat tttaaaaaat gctattgtat ataatagtat gtatgaaaat      240

tttagtacta gcttttggat aagaattcct aagtatttta acagtataag tctaaataat      300

gaatatacaa taataaattg tatggaaaat aattcaggat ggaaagtatc acttaattat      360

ggtgaaataa tctggacttt acaggatact caggaaataa aacaaagagt agtttttaaa      420

tacagtcaaa tgattaatat atcagattat ataaacagat ggatttttgt aactatcact      480

aataatagat taaataactc taaaatttat ataaatggaa gattaataga tcaaaaacca      540

atttcaaatt taggtaatat tcatgctagt aataatataa tgtttaaatt agatggttgt      600

agagatacac atagatatat ttggataaaa tattttaatc tttttgataa ggaattaaat      660

gaaaaagaaa tcaaagattt atatgataat caatcaaatt caggtatttt aaaagacttt      720

tggggtgatt atttacaata tgataaacca tactatatgt taaatttata tgatccaaat      780

aaatatgtcg atgtaaataa tgtaggtatt agaggttata tgtatcttaa agggcctaga      840

ggtagcgtaa tgactacaaa catttattta aattcaagtt tgtatagggg gacaaaattt      900

attataaaaa aatatgcttc tggaaataaa gataatattg ttagaaataa tgatcgtgta      960

tatattaatg tagtagttaa aaataaagaa tataggttag ctactaatgc atcacaggca     1020

ggcgtagaaa aaatactaag tgcattagaa atacctgatg taggaaatct aagtcaagta     1080

gtagtaatga agtcaaaaaa tgatcaagga ataacaaata aatgcaaaat gaatttacaa     1140

gataataatg ggaatgatat aggctttata ggatttcatc agtttaataa tatagctaaa     1200

ctagtagcaa gtaattggta taatagacaa atagaaagat ctagtaggac tttgggttgc     1260

tcatgggaat ttattcctgt agatgatgga tggggagaaa ggccactgta a              1311


<210>  5
<211>  1311
<212>  DNA
<213>  Artificial

<220>
<223>  codon-enhanced sequence encoding 50kD heavy chain fragment of 
       Clostridium botulinum neurotoxin A

<400>  5
cggctcctgt ccactttcac agaatatatc aaaaacatta tcaatactag catcctgaat       60

ctccggtatg agagcaacca cctgattgac ctgagcaggt acgcaagcaa aatcaacatc      120

ggctccaagg tgaactttga ccccatcgat aagaaccaaa ttcaactctt taatctcgaa      180

tccagcaaga ttgaggtcat tctgaaaaac gctatcgtgt acaactccat gtacgagaac      240

ttttccacca gcttctggat taggatccca aaatacttca atagcatttc cctcaataac      300

gagtatacca tcatcaattg tatggaaaac aatagcggct ggaaggtgtc cctgaattac      360

ggagagatca tctggactct gcaagacacc caggagatca aacagagagt cgtgttcaaa      420

tactcccaaa tgattaacat tagcgactac atcaacagat ggatcttcgt gacaattaca      480

aacaacaggc tgaataactc caagatttac attaacggca ggctcatcga tcagaagcct      540

attagcaacc tcggcaatat tcatgccagc aataacatca tgtttaagct cgacggctgt      600

agggataccc acaggtacat ttggatcaag tactttaacc tctttgacaa ggaactgaac      660

gaaaaagaaa tcaaagatct ctacgataac caaagcaact ccggaattct caaagacttt      720

tggggcgatt acctgcaata cgacaagcca tactacatgc tcaacctgta tgaccctaac      780

aaatacgtgg atgtcaataa tgtgggcatc cgggggtata tgtatctcaa gggacctcgg      840

ggaagcgtga tgaccacaaa tatctacctg aacagctccc tgtatagagg cactaaattc      900

atcattaaga agtatgctag cggaaacaag gataacattg tgcggaacaa cgacagagtc      960

tatattaacg tcgtcgtcaa gaacaaagag tacagactgg caacaaacgc atcccaggct     1020

ggagtcgaaa agattctcag cgctctggaa attcctgacg tgggcaatct cagccaggtc     1080

gtggtgatga aaagcaagaa cgatcagggc atcactaata agtgtaagat gaacctccag     1140

gacaacaacg ggaatgatat cggattcatt gggtttcacc agtttaacaa cattgctaaa     1200

ctcgtcgcct ccaactggta caataggcag atcgagagaa gcagcagaac actcgggtgt     1260

agctgggagt tcatccccgt cgatgacggg tggggagaac ggcctctctg a              1311


<210>  6
<211>  436
<212>  PRT
<213>  Artificial

<220>
<223>  50kD heavy chain fragment of Clostridium botulinum neurotoxin A

<400>  6

Arg Leu Leu Ser Thr Phe Thr Glu Tyr Ile Lys Asn Ile Ile Asn Thr 
1               5                   10                  15      


Ser Ile Leu Asn Leu Arg Tyr Glu Ser Asn His Leu Ile Asp Leu Ser 
            20                  25                  30          


Arg Tyr Ala Ser Lys Ile Asn Ile Gly Ser Lys Val Asn Phe Asp Pro 
        35                  40                  45              


Ile Asp Lys Asn Gln Ile Gln Leu Phe Asn Leu Glu Ser Ser Lys Ile 
    50                  55                  60                  


Glu Val Ile Leu Lys Asn Ala Ile Val Tyr Asn Ser Met Tyr Glu Asn 
65                  70                  75                  80  


Phe Ser Thr Ser Phe Trp Ile Arg Ile Pro Lys Tyr Phe Asn Ser Ile 
                85                  90                  95      


Ser Leu Asn Asn Glu Tyr Thr Ile Ile Asn Cys Met Glu Asn Asn Ser 
            100                 105                 110         


Gly Trp Lys Val Ser Leu Asn Tyr Gly Glu Ile Ile Trp Thr Leu Gln 
        115                 120                 125             


Asp Thr Gln Glu Ile Lys Gln Arg Val Val Phe Lys Tyr Ser Gln Met 
    130                 135                 140                 


Ile Asn Ile Ser Asp Tyr Ile Asn Arg Trp Ile Phe Val Thr Ile Thr 
145                 150                 155                 160 


Asn Asn Arg Leu Asn Asn Ser Lys Ile Tyr Ile Asn Gly Arg Leu Ile 
                165                 170                 175     


Asp Gln Lys Pro Ile Ser Asn Leu Gly Asn Ile His Ala Ser Asn Asn 
            180                 185                 190         


Ile Met Phe Lys Leu Asp Gly Cys Arg Asp Thr His Arg Tyr Ile Trp 
        195                 200                 205             


Ile Lys Tyr Phe Asn Leu Phe Asp Lys Glu Leu Asn Glu Lys Glu Ile 
    210                 215                 220                 


Lys Asp Leu Tyr Asp Asn Gln Ser Asn Ser Gly Ile Leu Lys Asp Phe 
225                 230                 235                 240 


Trp Gly Asp Tyr Leu Gln Tyr Asp Lys Pro Tyr Tyr Met Leu Asn Leu 
                245                 250                 255     


Tyr Asp Pro Asn Lys Tyr Val Asp Val Asn Asn Val Gly Ile Arg Gly 
            260                 265                 270         


Tyr Met Tyr Leu Lys Gly Pro Arg Gly Ser Val Met Thr Thr Asn Ile 
        275                 280                 285             


Tyr Leu Asn Ser Ser Leu Tyr Arg Gly Thr Lys Phe Ile Ile Lys Lys 
    290                 295                 300                 


Tyr Ala Ser Gly Asn Lys Asp Asn Ile Val Arg Asn Asn Asp Arg Val 
305                 310                 315                 320 


Tyr Ile Asn Val Val Val Lys Asn Lys Glu Tyr Arg Leu Ala Thr Asn 
                325                 330                 335     


Ala Ser Gln Ala Gly Val Glu Lys Ile Leu Ser Ala Leu Glu Ile Pro 
            340                 345                 350         


Asp Val Gly Asn Leu Ser Gln Val Val Val Met Lys Ser Lys Asn Asp 
        355                 360                 365             


Gln Gly Ile Thr Asn Lys Cys Lys Met Asn Leu Gln Asp Asn Asn Gly 
    370                 375                 380                 


Asn Asp Ile Gly Phe Ile Gly Phe His Gln Phe Asn Asn Ile Ala Lys 
385                 390                 395                 400 


Leu Val Ala Ser Asn Trp Tyr Asn Arg Gln Ile Glu Arg Ser Ser Arg 
                405                 410                 415     


Thr Leu Gly Cys Ser Trp Glu Phe Ile Pro Val Asp Asp Gly Trp Gly 
            420                 425                 430         


Glu Arg Pro Leu 
        435     


<210>  7
<211>  25
<212>  PRT
<213>  Artificial

<220>
<223>  25 residue signal peptide of human tissue plasminogen activator

<400>  7

Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly 
1               5                   10                  15      


Ala Val Phe Val Ser Pro Ser Gln Glu 
            20                  25  


<210>  8
<211>  81
<212>  DNA
<213>  Artificial

<220>
<223>  DNA sequence encoding signal peptide of human tissue plasminogen 
       activator and Ser-Ser linker

<400>  8
atggatgcaa tgaagagagg gctctgctgt gtgctgctgc tgtgtggagc agtcttcgtt       60

tcgcccagcc aggaaagcag c                                                 81


