                         SEQUENCE LISTING

<110>  University of Cape Town
 
<120>  Synthetic BTV VP2 Multiepitope Peptide Vaccine

<130>  PA159219/P

<160>  22    

<170>  PatentIn version 3.5

<210>  1
<211>  339
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Zera Signal Sequence

<400>  1
atgagggtgt tgctcgttgc cctcgctctc ctggctctcg ctgcgagcgc cacctccacg       60

catacaagcg gcggctgcgg ctgccagcca ccgccgccgg ttcatctacc gccgccggtg      120

catctgccac ctccggttca cctgccacct ccggtgcatc tcccaccgcc ggtccacctg      180

ccgccgccgg tccacctgcc accgccggtc catgtgccgc cgccggttca tctgccgccg      240

ccaccatgcc actaccctac tcaaccgccc cggcctcagc ctcatcccca gccacaccca      300

tgcccgtgcc aacagccgca tccaagcccg tgccagacc                             339


<210>  2
<211>  113
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Zera Signal Sequence

<400>  2

Met Arg Val Leu Leu Val Ala Leu Ala Leu Leu Ala Leu Ala Ala Ser 
1               5                   10                  15      


Ala Thr Ser Thr His Thr Ser Gly Gly Cys Gly Cys Gln Pro Pro Pro 
            20                  25                  30          


Pro Val His Leu Pro Pro Pro Val His Leu Pro Pro Pro Val His Leu 
        35                  40                  45              


Pro Pro Pro Val His Leu Pro Pro Pro Val His Leu Pro Pro Pro Val 
    50                  55                  60                  


His Leu Pro Pro Pro Val His Val Pro Pro Pro Val His Leu Pro Pro 
65                  70                  75                  80  


Pro Pro Cys His Tyr Pro Thr Gln Pro Pro Arg Pro Gln Pro His Pro 
                85                  90                  95      


Gln Pro His Pro Cys Pro Cys Gln Gln Pro His Pro Ser Pro Cys Gln 
            100                 105                 110         


Thr 
    


<210>  3
<211>  18
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  VP2 Epitope

<400>  3

Phe Pro Glu His Leu Arg His Asp Val Thr Asn Pro Tyr Pro Cys Leu 
1               5                   10                  15      


Arg Gly 
        


<210>  4
<211>  115
<212>  PRT
<213>  Bluetongue virus

<400>  4

Met Asp Gly Leu Gly Ile Pro Ile Tyr Lys Gln Gly Phe Pro Glu His 
1               5                   10                  15      


Leu Leu His Gly Tyr Glu Phe Thr Ile Asp Ser Ser Thr Lys Ile Gln 
            20                  25                  30          


Ser Val Gly Gly Arg His Asp Val Thr Lys Leu Pro Glu Met Asn Ala 
        35                  40                  45              


Tyr Asp Ile Lys Ala Glu Ser Ile Trp Trp Ser Asn Pro Tyr Pro Cys 
    50                  55                  60                  


Leu Arg Gly Thr Leu Met Ala Ser Glu Thr Lys Leu Gly Asp Val Tyr 
65                  70                  75                  80  


Ser Met Ile Arg Ser Trp Tyr Asp Trp Ser Val Arg Pro Thr Tyr Thr 
                85                  90                  95      


Pro Tyr Glu Lys Ser Arg Glu Gln Glu Lys Tyr Ile Tyr Gly Arg Val 
            100                 105                 110         


Asn Leu Phe 
        115 


<210>  5
<211>  115
<212>  PRT
<213>  Bluetonge virus

<400>  5

Met Asp Glu Leu Gly Ile Pro Ile Tyr Gly Arg Asn Tyr Pro Glu His 
1               5                   10                  15      


Leu Leu Lys Gly Tyr Glu Phe Leu Ile Asn Thr Gly Val Arg Tyr Pro 
            20                  25                  30          


Ser Gln Gly Gly Arg His Asp Val Ser Lys Ile Pro Glu Met Phe Ala 
        35                  40                  45              


Tyr Asp Ile Lys Asp Glu Gly Ile Trp Trp Ser Asn Pro Tyr Pro Cys 
    50                  55                  60                  


Leu Arg Gly Thr Leu Ile Ala Ala Glu Thr Lys Leu Gly Asp Val Tyr 
65                  70                  75                  80  


Phe Thr Leu Arg Ser Trp Tyr Asp Trp Ser Val Arg Ser Ser Tyr Val 
                85                  90                  95      


Pro Arg Glu Arg Glu Arg Glu Thr Glu Lys Tyr Ile Phe Ser Lys Ile 
            100                 105                 110         


Asn Leu Phe 
        115 


<210>  6
<211>  113
<212>  PRT
<213>  Bluetonge virus

<400>  6

Met Asp Glu Leu Ala Ile Pro Ile Ile Asn Gly His Phe Pro Lys Glu 
1               5                   10                  15      


Ala Leu Ile Glu Tyr Asp Cys Ile Val Glu Leu Asn Ala Pro Ser Glu 
            20                  25                  30          


Asp Asp Glu Val Asp Val Thr Gln Ile Pro Thr Lys Asn Met Met Asp 
        35                  40                  45              


Ile Pro Asp Val Pro Ile Trp Trp Thr Asn Pro Tyr Pro Cys Leu Arg 
    50                  55                  60                  


Gly Ala Leu Ile Ala Ser Glu Thr Arg Leu Gly Asp Val Tyr Trp Thr 
65                  70                  75                  80  


Leu Arg His Thr Tyr Asp Trp Ser Val Arg Met Thr Tyr Ser Pro Arg 
                85                  90                  95      


Met Lys Glu Arg Glu Asp Asp Lys Tyr Ile Tyr Gly Arg Val Asn Leu 
            100                 105                 110         


Phe 
    


<210>  7
<211>  119
<212>  PRT
<213>  Bluetonge virus

<400>  7

Met Glu Glu Leu Ala Ile Pro Ile Tyr Thr Asn Val Phe Pro Ala Glu 
1               5                   10                  15      


Leu Leu Asp Gly Tyr Asp Tyr Ile Ile Asp Val Ser Ser Arg Val Glu 
            20                  25                  30          


Glu Glu Gly Asp Glu Pro Val Lys Arg His Asp Val Thr Glu Ile Pro 
        35                  40                  45              


Arg Asn Ser Met Phe Asp Ile Lys Asp Glu His Ile Trp Trp Ser Asn 
    50                  55                  60                  


Pro Tyr Pro Cys Leu Arg Gly Ala Leu Ile Ala Ala Glu Val Gln Leu 
65                  70                  75                  80  


Gly Asp Val Tyr Asn Leu Leu Arg Asn Trp Phe Gln Trp Ser Val Arg 
                85                  90                  95      


Pro Thr Tyr Val Pro Tyr Asp Arg Asn Arg Glu Ser Asp Lys Tyr Ile 
            100                 105                 110         


Tyr Ser Arg Ile Asn Leu Phe 
        115                 


<210>  8
<211>  116
<212>  PRT
<213>  Bluetonge virus

<400>  8

Met Glu Glu Phe Val Ile Pro Val Phe Ser Glu Arg Asp Ile Pro Tyr 
1               5                   10                  15      


Leu Leu Leu Asn His Tyr Pro Leu Ala Ile Gln Ile Asp Val Lys Val 
            20                  25                  30          


Asp Asp Glu Gly Gly Lys His Asn Leu Ile Lys Ile Pro Glu Ser Asp 
        35                  40                  45              


Met Ile Asp Val Pro Arg Leu Ser Val Trp Arg Thr Asn Pro Tyr Pro 
    50                  55                  60                  


Cys Leu Arg Gly Ala Leu Ile Ala Ala Glu Cys Glu Leu Gly Asp Val 
65                  70                  75                  80  


Tyr Phe Thr Leu Arg Gln Thr Tyr Lys Trp Ser Leu Arg Pro Glu Tyr 
                85                  90                  95      


Gly Gln Arg Glu Arg Pro Leu Glu Asp Asn Lys Tyr Val Phe Ala Arg 
            100                 105                 110         


Leu Asn Leu Phe 
        115     


<210>  9
<211>  115
<212>  PRT
<213>  Bluetonge virus

<400>  9

Met Glu Glu Leu Val Ile Pro Val Ile Thr Glu Arg Phe Asp Lys Arg 
1               5                   10                  15      


Leu Val Val Arg Tyr Asp Tyr Val Ile Glu Leu Ser Ser Pro Glu Gly 
            20                  25                  30          


Asp Glu Trp Asn Gly His Asp Val Thr His Ile Pro Asp Lys Arg Met 
        35                  40                  45              


Phe Asp Ile Lys Val Gln Pro Val Trp Trp Ser Asn Pro Tyr Pro Cys 
    50                  55                  60                  


Leu Arg Gly Ala Leu Ile Ala Ala Glu Cys Lys Leu Gly Asp Val Tyr 
65                  70                  75                  80  


Tyr Lys Leu Arg Ser Trp Tyr Glu Trp Ser Val Arg Glu Gly Tyr Lys 
                85                  90                  95      


Pro Arg Asp Leu Asp Arg Gln Tyr Glu Lys Tyr Ile Val Gly Arg Val 
            100                 105                 110         


Asn Leu Phe 
        115 


<210>  10
<211>  116
<212>  PRT
<213>  Bluetonge virus

<400>  10

Met Glu Glu Phe Val Ile Pro Val Tyr Ser Glu Asp Glu Ile Pro Tyr 
1               5                   10                  15      


Ala Leu Leu Ser Arg Tyr Pro Leu Ala Ile Gln Thr Asn Val Lys Ile 
            20                  25                  30          


Glu Asp Val Glu Gly Arg His Asn Val Val Glu Ile Pro Glu Ser Asp 
        35                  40                  45              


Met Ile Asp Ile Pro Arg Leu Arg Val Trp Arg Thr Asn Pro Tyr Pro 
    50                  55                  60                  


Cys Leu Arg Gly Ala Leu Val Ala Ala Glu Cys Glu Leu Gly Asp Val 
65                  70                  75                  80  


Tyr Ser Thr Leu Arg Arg Val Tyr Arg Trp Ser Leu Arg Pro Glu Tyr 
                85                  90                  95      


Gly Gln His Glu Arg Gln Leu Glu Asn Asn Lys Tyr Val Phe Asn Arg 
            100                 105                 110         


Ile Asn Leu Phe 
        115     


<210>  11
<211>  115
<212>  PRT
<213>  Bluetonge virus

<400>  11

Met Asp Glu Leu Ala Ile Pro Ile Tyr Thr Asn Val Phe Pro Ala Glu 
1               5                   10                  15      


Leu Leu Asp Gly Tyr Asp Tyr Ile Ile Asp Val Ser Thr Arg Ile Asp 
            20                  25                  30          


Glu Pro Ala Gly Arg His Asp Val Thr Gln Ile Pro Asn Ser Ala Met 
        35                  40                  45              


Phe Asp Val Gln Asn Glu His Ile Trp Trp Ser Asn Pro Tyr Pro Cys 
    50                  55                  60                  


Leu Arg Gly Ser Leu Ile Ala Ala Glu Ala Gln Leu Gly Asp Val Tyr 
65                  70                  75                  80  


Asn Thr Met Arg Asn Trp Tyr Gln Trp Ser Val Arg Pro Thr Tyr Thr 
                85                  90                  95      


Pro Tyr Asp Arg Ser Arg Glu Ser Asp Lys Tyr Val Tyr Ser Arg Val 
            100                 105                 110         


Asn Leu Phe 
        115 


<210>  12
<211>  54
<212>  DNA
<213>  Artificial sequence

<220>
<223>  BTV VP2 epitope

<400>  12
ttcccagagc accttcgaca cgatgtaaca aatccgtatc cctgcctgcg tgga             54


<210>  13
<211>  41
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Synthesized oligonucleotide for assembly of BTV VP2 epitope 
       construct containing MluI restriction site

<400>  13
gcacgcgttt cccagagcac cttcgacacg atgtaacaaa t                           41


<210>  14
<211>  41
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Synthesized oligonucleotide for assembly of BTV VP2 epitope 
       construct containing XmaI restriction site

<400>  14
ctacattgtt taggcatagg gacggcagca cctgggcccg c                           41


<210>  15
<211>  4
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Universal alpha-helix linker

<400>  15

Glu Ala Ala Lys 
1               


<210>  16
<211>  27
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Universal alpha-helix linker flanked by NcoI and MluI restriction
       sites

<400>  16
ccatgggaag cggcggcgaa aacgcgt                                           27


<210>  17
<211>  26
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Oligonucleotide - Zera FP

<400>  17
gcaccggtat gagggtgttg ctcgtt                                            26


<210>  18
<211>  25
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Oligonucleotide - Zera-RP

<400>  18
gcccatggct ggcacgggct tggat                                             25


<210>  19
<211>  432
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Zera-Linker-BTV VP2 epitope

<400>  19
gcaccggtat gagggtgttg ctcgttgccc tcgctctcct ggctctcgct gcgagcgcca       60

cctccacgca tacaagcggc ggctgcggct gccagccacc gccgccggtt catctaccgc      120

cgccggtgca tctgccacct ccggttcacc tgccacctcc ggtgcatctc ccaccgccgg      180

tccacctgcc gccgccggtc cacctgccac cgccggtcca tgtgccgccg ccggttcatc      240

tgccgccgcc accatgccac taccctactc aaccgccccg gcctcagcct catccccagc      300

cacacccatg cccgtgccaa cagccgcatc caagcccgtg ccagaccatg ggaagcggcg      360

gcgaaaacgc gtttcccaga gcaccttcga cacgatgtaa caaatccgta tccctgccgt      420

cgtggacccg gg                                                          432


<210>  20
<211>  147
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Zera-Linker-BTV VP2 epitope

<400>  20

Met Arg Val Leu Leu Val Ala Leu Ala Leu Leu Ala Leu Ala Ala Ser 
1               5                   10                  15      


Ala Thr Ser Thr His Thr Ser Gly Gly Cys Gly Cys Gln Pro Pro Pro 
            20                  25                  30          


Pro Val His Leu Pro Pro Pro Val His Leu Pro Pro Pro Val His Leu 
        35                  40                  45              


Pro Pro Pro Val His Leu Pro Pro Pro Val His Leu Pro Pro Pro Val 
    50                  55                  60                  


His Leu Pro Pro Pro Val His Val Pro Pro Pro Val His Leu Pro Pro 
65                  70                  75                  80  


Pro Pro Cys His Tyr Pro Thr Gln Pro Pro Arg Pro Gln Pro His Pro 
                85                  90                  95      


Gln Pro His Pro Cys Pro Cys Gln Gln Pro His Pro Ser Pro Cys Gln 
            100                 105                 110         


Pro Trp Glu Ala Ala Ala Lys Thr Arg Phe Pro Glu His Leu Arg His 
        115                 120                 125             


Asp Val Thr Asn Pro Tyr Pro Cys Leu Arg Gly Pro Gly His His His 
    130                 135                 140                 


His His His 
145         


<210>  21
<211>  1110
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  BTV VP2 Multiepitope

<400>  21
atgagggtgt tgctcgttgc cctcgctctc ctggctctcg ctgcgagcgc cacctccacg       60

catacaagcg gcggctgcgg ctgccagcca ccgccgccgg ttcatctacc gccgccggtg      120

catctgccac ctccggttca cctgccacct ccggtgcatc tcccaccgcc ggtccacctg      180

ccgccgccgg tccacctgcc accgccggtc catgtgccgc cgccggttca tctgccgccg      240

ccaccatgcc actaccctac tcaaccgccc cggcctcagc ctcatcccca gccacaccca      300

tgcccgtgcc aacagccgca tccaagcccg tgccagacca tggaagctgc tgctaagacc      360

ggtatgggtc ctggtcctgg tcaatttgat aaaaaactcg gtcctggtcc tggagaactt      420

gctgagcctg aagatgatga gtggagtgga catgatgtta ctcaaattcc tgatggtcca      480

ggtccaggag atattaagga tgaatccggt ccaggacctg gagatgtgcc taatggtgaa      540

ggacctggac ctggagatgt tccaaaggtg gatggtcctg gtcctggatc tggttatgat      600

agaaggaaat accacaagac aaattctgga ggtgaatttt attcaccagg acctggtcca      660

ggagaaaaca ctgagaagat gaatatcatg caaaacaagt caggaagaac agatgatgag      720

aggggtcctg gaccaggtac tgatacacag aggggtaggg tttggaggat gaatccttac      780

ccatgtctta ggggaactgg tcctggacca ggtgttagac ctacatatgt gccatacgaa      840

agagagaggg aaactgagaa atatatatac agtggacctg gtcctggaaa gtttaataag      900

atggctggtc ctggaccagg ttgggatcaa gaaagattca aaggtccagg acctggaaac      960

tttacaaggt tccatccagg agattcaaag aaaaagcaga aggcagatga tactcacggt     1020

cctggaccag gttattacac aaatatgaaa atctctcaag gtggtgtgaa tgtgcctgtg     1080

gtgacttcca agggtcctgg tccaggatga                                      1110


<210>  22
<211>  369
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  BTV VP2 Multiepitope

<400>  22

Met Arg Val Leu Leu Val Ala Leu Ala Leu Leu Ala Leu Ala Ala Ser 
1               5                   10                  15      


Ala Thr Ser Thr His Thr Ser Gly Gly Cys Gly Cys Gln Pro Pro Pro 
            20                  25                  30          


Pro Val His Leu Pro Pro Pro Val His Leu Pro Pro Pro Val His Leu 
        35                  40                  45              


Pro Pro Pro Val His Leu Pro Pro Pro Val His Leu Pro Pro Pro Val 
    50                  55                  60                  


His Leu Pro Pro Pro Val His Val Pro Pro Pro Val His Leu Pro Pro 
65                  70                  75                  80  


Pro Pro Cys His Tyr Pro Thr Gln Pro Pro Arg Pro Gln Pro His Pro 
                85                  90                  95      


Gln Pro His Pro Cys Pro Cys Gln Gln Pro His Pro Ser Pro Cys Gln 
            100                 105                 110         


Thr Met Glu Ala Ala Ala Lys Thr Gly Met Gly Pro Gly Pro Gly Gln 
        115                 120                 125             


Phe Asp Lys Lys Leu Gly Pro Gly Pro Gly Glu Leu Ala Glu Pro Glu 
    130                 135                 140                 


Asp Asp Glu Trp Ser Gly His Asp Val Thr Gln Ile Pro Asp Gly Pro 
145                 150                 155                 160 


Gly Pro Gly Asp Ile Lys Asp Glu Ser Gly Pro Gly Pro Gly Asp Val 
                165                 170                 175     


Pro Asn Gly Glu Gly Pro Gly Pro Gly Asp Val Pro Lys Val Asp Gly 
            180                 185                 190         


Pro Gly Pro Gly Ser Gly Tyr Asp Arg Arg Lys Tyr His Lys Thr Asn 
        195                 200                 205             


Ser Gly Gly Glu Phe Tyr Ser Pro Gly Pro Gly Pro Gly Glu Asn Thr 
    210                 215                 220                 


Glu Lys Met Asn Ile Met Gln Asn Lys Ser Gly Arg Thr Asp Asp Glu 
225                 230                 235                 240 


Arg Gly Pro Gly Pro Gly Thr Asp Thr Gln Arg Gly Arg Val Trp Arg 
                245                 250                 255     


Met Asn Pro Tyr Pro Cys Leu Arg Gly Thr Gly Pro Gly Pro Gly Val 
            260                 265                 270         


Arg Pro Thr Tyr Val Pro Tyr Glu Arg Glu Arg Glu Thr Glu Lys Tyr 
        275                 280                 285             


Ile Tyr Ser Gly Pro Gly Pro Gly Lys Phe Asn Lys Met Ala Gly Pro 
    290                 295                 300                 


Gly Pro Gly Trp Asp Gln Glu Arg Phe Lys Gly Pro Gly Pro Gly Asn 
305                 310                 315                 320 


Phe Thr Arg Phe His Pro Gly Asp Ser Lys Lys Lys Gln Lys Ala Asp 
                325                 330                 335     


Asp Thr His Gly Pro Gly Pro Gly Tyr Tyr Thr Asn Met Lys Ile Ser 
            340                 345                 350         


Gln Gly Gly Val Asn Val Pro Val Val Thr Ser Lys Gly Pro Gly Pro 
        355                 360                 365             


Gly 
    


