                         SEQUENCE LISTING

<110>  Hughes, Kelly
       Olivera, Baldomero
 
<120>  COMPOSITIONS AND METHODS FOR PEPTIDE EXPRESSION AND PURIFICATION 
       USING A TYPE III SECRETION SYSTEM

<130>  21101.0290P1

<150>  61/689,284
<151>  2012-05-30

<160>  73    

<170>  PatentIn version 3.5

<210>  1
<211>  66
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  1
cagaactgct gcaacggcgg ctgcagcagc aaatggtgcc gcgatcatgc gcgctgctgc       60

ggccgc                                                                  66


<210>  2
<211>  22
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA peptide

<400>  2

Gln Asn Cys Cys Asn Gly Gly Cys Ser Ser Lys Trp Cys Arg Asp His 
1               5                   10                  15      


Ala Arg Cys Cys Gly Arg 
            20          


<210>  3
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  3
gcggccgcag cagcgcgcat g                                                 21


<210>  4
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  synthetic construct; TEV protease cleavage site

<400>  4

Glu Asn Leu Tyr Phe Gln Gly 
1               5           


<210>  5
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; histidine tag

<400>  5
catcaccatc accatcac                                                     18


<210>  6
<211>  51
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; FlgM primer

<400>  6
cgggatcccg atgcatcacc atcaccatca catgagcatt gaccgtacct c                51


<210>  7
<211>  59
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; FlgM primer

<400>  7
ccgttgcagc agttctggcc ctgaaaatac aggttttctt tactctgtaa gtagctctg        59


<210>  8
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  8
ttttcagggc cagaactgct gcaacggcgg                                        30


<210>  9
<211>  28
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  9
ggaattcctt agcggccgca gcagcgcg                                          28


<210>  10
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; FlgM primer

<400>  10
actcgctcat tcgcgaggcg cagagctact tacagagtaa aggcagctct caccaccacc       60


<210>  11
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; FlgM primer

<400>  11
ttcatcaacg cgccccccat gggacgcgtt tttagaggca ttaacggtta cctgcacaag       60


<210>  12
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; FlgM primer

<400>  12
cgggatcccg atgagcattg accgtacctc                                        30


<210>  13
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; FlgM primer

<400>  13
ccgttgcagc agttctggcc ctgaaaatac aggttttctt tactctgtaa gtagctctgc       60


<210>  14
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  14
ttcagggcca gaactgctgc aacggc                                            26


<210>  15
<211>  49
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  15
ggaattcctt agtgatggtg atggtgatgg cggccgcagc agcgcgcat                   49


<210>  16
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; FlgM primer

<400>  16
gccctgaaaa tacaggtttt cgtgatggtg atggtgatgt ttactctgta agtagctctg       60


<210>  17
<211>  46
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  17
tcacgaaaac ctgtattttc agggccagaa ctgctgcaac ggcggc                      46


<210>  18
<211>  28
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  18
ggaattcctt agcggccgca gcagcgcg                                          28


<210>  19
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; MVIIA primer

<400>  19
tgcaaaggta aaggtgcaaa atgtagccgt ctgatgtatg attgttgtac cggtagctgt       60


<210>  20
<211>  38
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; MVIIA primer

<400>  20
ttaacattta ccgctacgac agctaccggt acaacaat                               38


<210>  21
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; MVIIA primer

<400>  21
acatcaccat caccatcacg aaaacctgta ttttcagggc tgcaaaggta aaggtgcaaa       60


<210>  22
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; MVIIA primer

<400>  22
ttcatcaacg cgccccccat gggacgcgtt tttagaggca ttaacattta ccgctacgac       60


<210>  23
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; GVIA primer

<400>  23
tgtaaaagtc cgggtagcag ctgtagcccg accagctata attgttgtcg tagctgtaat       60


<210>  24
<211>  44
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; GVIA primer

<400>  24
ttaatagcaa cgtttggtat acggattaca gctacgacaa caat                        44


<210>  25
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; GVIA primer

<400>  25
acatcaccat caccatcacg aaaacctgta ttttcagggc tgtaaaagtc cgggtagcag       60


<210>  26
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; GVIA primer

<400>  26
ttcatcaacg cgccccccat gggacgcgtt tttagaggca ttaatagcaa cgtttggtat       60


<210>  27
<211>  51
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; contulakin primer

<400>  27
gaaagcgaag aaggtggtag caacgcaacc aaaaaaccgt atattctgta a                51


<210>  28
<211>  51
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; contulakin primer

<400>  28
ttacagaata tacggttttt tggttgcgtt gctaccacct tcttcgcttt c                51


<210>  29
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; contulakin primer

<400>  29
acatcaccat caccatcacg aaaacctgta ttttcagggc gaaagcgaag aaggtggtag       60


<210>  30
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; contulakin primer

<400>  30
ttcatcaacg cgccccccat gggacgcgtt tttagaggca ttacagaata tacggttttt       60


<210>  31
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; alpha Vc1.1 primer

<400>  31
tgttgtagcg atccgcgttg taattatgat catccggaaa tttgctaa                    48


<210>  32
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; alpha Vc1.1 primer

<400>  32
ttagcaaatt tccggatgat cataattaca acgcggatcg ctacaaca                    48


<210>  33
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; alpha Vc1.1 primer

<400>  33
acatcaccat caccatcacg aaaacctgta ttttcagggc tgttgtagcg atccgcgttg       60


<210>  34
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; alpha Vc1.1 primer

<400>  34
ttcatcaacg cgccccccat gggacgcgtt tttagaggca ttagcaaatt tccggatgat       60


<210>  35
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; GsMTx4 primer

<400>  35
tgtctggaat tttggtggaa atgcaatccg aacgatgata aatgttgtcg tccgaaactg       60


<210>  36
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; GsMTx4 primer

<400>  36
accgctgcta aaattgcaca gtttaaacag tttgctgcat ttcagtttcg gacgacaaca       60


<210>  37
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; GsMTx4 primer

<400>  37
acatcaccat caccatcacg aaaacctgta ttttcagggc tgtctggaat tttggtggaa       60


<210>  38
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; GsMTx4 primer

<400>  38
atcaacgcgc cccccatggg acgcgttttt agaggcatta accgctgcta aaattgcaca       60


<210>  39
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Calciseptine primer

<400>  39
cgcatctgct atattcataa agcaagcctg cctcgtgcaa ccaaaacctg tgttgaaaat       60


<210>  40
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Calciseptine primer

<400>  40
tatattcgcg ctgggtacga ataaacattt tatagcaggt attttcaaca caggttttgg       60


<210>  41
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Calciseptine primer

<400>  41
tcgtacccag cgcgaatata tcagcgaacg tggttgtggt tgtccgaccg caatgtggcc       60


<210>  42
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Calciseptine primer

<400>  42
tttgttgcaa cgatcacctt tacaacattc ggtctgatac ggccacattg cggtcggaca       60


<210>  43
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Calciseptine primer

<400>  43
acatcaccat caccatcacg aaaacctgta ttttcagggc cgcatctgct atattcataa       60


<210>  44
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Calciseptine primer

<400>  44
atcaacgcgc cccccatggg acgcgttttt agaggcatta tttgttgcaa cgatcacctt       60


<210>  45
<211>  54
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Conantokin primer

<400>  45
ggtgaagaag aactgcaaga aaaccaagaa ctgattcgcg aaaaaagcaa ttaa             54


<210>  46
<211>  54
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Conantokin primer

<400>  46
ttaattgctt ttttcgcgaa tcagttcttg gttttcttgc agttcttctt cacc             54


<210>  47
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Conantokin primer

<400>  47
acatcaccat caccatcacg aaaacctgta ttttcagggc ggtgaagaag aactgcaaga       60


<210>  48
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Conantokin primer

<400>  48
ttcatcaacg cgccccccat gggacgcgtt tttagaggca ttaattgctt ttttcgcgaa       60


<210>  49
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  49
gaaaattgct gtaatggtgg ttgtagcagc aaatggtgtc gtgatcatgc acgttgttgt       60


<210>  50
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  50
ttaacaacaa cgtgcatgat cacgacacca tttgctgcta caaccaccat tacagcaatt       60


<210>  51
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  51
acatcaccat caccatcacg aaaacctgta ttttcagggc gaaaattgct gtaatggtgg       60


<210>  52
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; SIIIA primer

<400>  52
ttcatcaacg cgccccccat gggacgcgtt tttagaggca ttaacaacaa cgtgcatgat       60


<210>  53
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Shk primer

<400>  53
agctgcattg ataccattcc gaaaagccgt tgtaccgcat ttcagtgtaa acacagcatg       60


<210>  54
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Shk primer

<400>  54
acaggtgcca caggttttac gacaaaagct cagacgatat ttcatgctgt gtttacactg       60


<210>  55
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Shk primer

<400>  55
atcaccatca ccatcacgaa aacctgtatt ttcagggccg tagctgcatt gataccattc       60


<210>  56
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Shk primer

<400>  56
catcaacgcg ccccccatgg gacgcgtttt tagaggcatt aacaggtgcc acaggtttta       60


<210>  57
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Chlorotoxin primer

<400>  57
atgtgtatgc cgtgttttac caccgatcat cagatggcac gtaaatgtga tgattgttgt       60


<210>  58
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Chlorotoxin primer

<400>  58
gacactgcgg accataacat ttaccgcgac ctttaccacc acaacaatca tcacatttac       60


<210>  59
<211>  31
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Chlorotoxin primer

<400>  59
ttaacgacac agacactgcg gaccataaca t                                      31


<210>  60
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Chlorotoxin primer

<400>  60
acatcaccat caccatcacg aaaacctgta ttttcagggc atgtgtatgc cgtgttttac       60


<210>  61
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic construct; Chlorotoxin primer

<400>  61
ttcatcaacg cgccccccat gggacgcgtt tttagaggca ttaacgacac agacactgcg       60


<210>  62
<211>  25
<212>  PRT
<213>  Conus magus

<400>  62

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


Thr Gly Ser Cys Arg Ser Gly Lys Cys 
            20                  25  


<210>  63
<211>  27
<212>  PRT
<213>  Conus geographus

<400>  63

Cys Lys Ser Pro Gly Ser Ser Cys Ser Pro Thr Ser Tyr Asn Cys Cys 
1               5                   10                  15      


Arg Ser Cys Asn Pro Tyr Thr Lys Arg Cys Tyr 
            20                  25          


<210>  64
<211>  16
<212>  PRT
<213>  Conus geographus

<400>  64

Glu Ser Glu Glu Gly Gly Ser Asn Ala Thr Lys Lys Pro Tyr Ile Leu 
1               5                   10                  15      


<210>  65
<211>  16
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  synthetic construct; conus victoriae alpha Vc1.1

<400>  65

Gly Cys Cys Ser Asp Pro Arg Cys Asn Tyr Asp His Pro Glu Ile Cys 
1               5                   10                  15      


<210>  66
<211>  17
<212>  PRT
<213>  Conus geographus

<400>  66

Gly Glu Glu Glu Leu Gln Glu Asn Gln Glu Leu Ile Arg Glu Lys Ser 
1               5                   10                  15      


Asn 
    


<210>  67
<211>  20
<212>  PRT
<213>  Conus striatus

<400>  67

Glu Asn Cys Cys Asn Gly Gly Cys Ser Ser Lys Trp Cys Arg Asp His 
1               5                   10                  15      


Ala Arg Cys Cys 
            20  


<210>  68
<211>  22
<212>  PRT
<213>  Conus striatus

<400>  68

Gln Asn Cys Cys Asn Gly Gly Cys Ser Ser Lys Trp Cys Arg Asp His 
1               5                   10                  15      


Ala Arg Cys Cys Gly Arg 
            20          


<210>  69
<211>  35
<212>  PRT
<213>  Stichodactyla helianthus

<400>  69

Arg Ser Cys Ile Asp Thr Ile Pro Lys Ser Arg Cys Thr Ala Phe Gln 
1               5                   10                  15      


Cys Lys His Ser Met Lys Tyr Arg Leu Ser Phe Cys Arg Lys Thr Cys 
            20                  25                  30          


Gly Thr Cys 
        35  


<210>  70
<211>  36
<212>  PRT
<213>  Leiurus quinquestriatus

<400>  70

Met Cys Met Pro Cys Phe Thr Thr Asp His Gln Met Ala Arg Lys Cys 
1               5                   10                  15      


Asp Asp Cys Cys Gly Gly Lys Gly Arg Gly Lys Cys Tyr Gly Pro Gln 
            20                  25                  30          


Cys Leu Cys Arg 
        35      


<210>  71
<211>  35
<212>  PRT
<213>  Grammostola spatulata

<400>  71

Gly Cys Leu Glu Phe Trp Trp Lys Cys Asn Pro Asn Asp Asp Lys Cys 
1               5                   10                  15      


Cys Arg Pro Lys Leu Lys Cys Ser Lys Leu Phe Lys Leu Cys Asn Phe 
            20                  25                  30          


Ser Ser Gly 
        35  


<210>  72
<211>  60
<212>  PRT
<213>  Dendroaspis polylepis

<400>  72

Arg Ile Cys Tyr Ile His Lys Ala Ser Leu Pro Arg Ala Thr Lys Thr 
1               5                   10                  15      


Cys Val Glu Asn Thr Cys Tyr Lys Met Phe Ile Arg Thr Gln Arg Glu 
            20                  25                  30          


Tyr Ile Ser Glu Arg Gly Cys Gly Cys Pro Thr Ala Met Trp Pro Tyr 
        35                  40                  45              


Gln Thr Glu Cys Cys Lys Gly Asp Arg Cys Asn Lys 
    50                  55                  60  


<210>  73
<211>  190
<212>  PRT
<213>  Corynebacterium diphtheriae

<400>  73

Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met Glu Asn 
1               5                   10                  15      


Phe Ser Ser Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser Ile Gln 
            20                  25                  30          


Lys Gly Ile Gln Lys Pro Lys Ser Gly Thr Gln Gly Asn Tyr Asp Asp 
        35                  40                  45              


Asp Trp Lys Gly Phe Tyr Ser Thr Asp Asn Lys Tyr Asp Ala Ala Gly 
    50                  55                  60                  


Ala Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly Gly Val 
65                  70                  75                  80  


Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu Lys Val 
                85                  90                  95      


Asp Asn Ala Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu Thr Glu 
            100                 105                 110         


Pro Leu Met Glu Gln Val Gly Thr Glu Glu Phe Ile Lys Arg Phe Gly 
        115                 120                 125             


Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro Phe Ala Glu Gly Ser 
    130                 135                 140                 


Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala Leu Ser 
145                 150                 155                 160 


Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly Gln Asp 
                165                 170                 175     


Ala Met Tyr Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg 
            180                 185                 190 


