                                SEQUENCE LISTING

<110> Gonzalez-Sapienza, Gualberto
      Hammock, Bruce
      Gonzalez-Techera, Andres
      Vanrell, Lucia
      Lassabe, Gabriel
      The Regents of the University of California
      Universidad de la Republica

<120> Non-Competitive Immunoassays to Detect
  Small Molecules Using Nanopeptamers
  

<130> 76916-893464

<140> WO Not yet assigned 
<141> Not yet assigned  

<150> US 61/732,524       
<151> 2012-12-03  

<160> 32

<170> FastSEQ for Windows Version 4.0

<210> 1
<211> 14
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic immune complex specific biotinylated peptide A
      (pA), biotinylated anti-molinate IC peptide, biotinylated
      peptide A specific for anti-molinate monoclonal antibody
      MoAb 14D7-molinate immune complex (IC)

<221> MOD_RES        
<222> (1)...(1)
<223> Ser modified by biotin

<221> PEPTIDE        
<222> (1)...(4)
<223> spacer between biotin and anti-IC peptide

<221> PEPTIDE        
<222> (5)...(14)
<223> peptide A

<400> 1
Ser Gly Ser Gly Cys Ser Leu Trp Asp Thr Thr Gly Trp Cys
 1               5                  10                  


<210> 2
<211> 14
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic biotinylated immune complex specific peptide 1M
      (p1M), biotinylated anti-molinate IC peptide, biotinylated
      peptide 1M specific for anti-molinate monoclonal antibody
      MoAb 14D7-molinate immune complex (IC)

<220> 
<221> MOD_RES        
<222> (1)...(1)
<223> Ser modified by biotin

<220> 
<221> PEPTIDE        
<222> (1)...(4)
<223> spacer between biotin and anti-IC peptide

<220> 
<221> PEPTIDE        
<222> (5)...(14)
<223> peptide 1M

<400> 2
Ser Gly Ser Gly Cys Ser Thr Trp Asp Thr Thr Gly Trp Cys
 1               5                  10                  


<210> 3
<211> 14
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic biotinylated immune complex specific peptide ICX11
      (pICX11), biotinylated anti-clomazone IC peptide, biotinylated
      peptide ICX11 specific for anti-clomazone monoclonal
      antibody MoAb 5.6-clomazone immune complex (IC)

<220> 
<221> MOD_RES        
<222> (1)...(1)
<223> Ser modified by biotin

<220> 
<221> PEPTIDE        
<222> (1)...(4)
<223> spacer between biotin and anti-IC peptide

<220> 
<221> PEPTIDE        
<222> (5)...(14)
<223> peptide ICX11

<400> 3
Ser Gly Ser Gly Cys Leu Glu Ala Pro Asn Ile Glu Gly Cys
 1               5                  10                  


<210> 4
<211> 10
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic immune complex specific peptide A (pA),
      anti-molinate IC peptide, peptide A specific for
      anti-molinate monoclonal antibody MoAb 14D7-molinate
      immune complex (IC)

<400> 4
Cys Ser Leu Trp Asp Thr Thr Gly Trp Cys
 1               5                  10  


<210> 5
<211> 10
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic immune complex specific peptide 1M (p1M),
      anti-molinate IC peptide, peptide 1M specific for
      anti-molinate monoclonal antibody MoAb 14D7-molinate
      immune complex (IC)

<400> 5
Cys Ser Thr Trp Asp Thr Thr Gly Trp Cys
 1               5                  10  


<210> 6
<211> 10
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic immune complex specific peptide ICX11
      (pICX11), anti-clomazone IC peptide, peptide ICX11
      specific for anti-clomazone monoclonal antibody
      MoAb 5.6-clomazone immune complex (IC)

<400> 6
Cys Leu Glu Ala Pro Asn Ile Glu Gly Cys
 1               5                  10  


<210> 7
<211> 10
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic peptide A9 (pA9), anti-atrazine IC
      peptide, peptide A9 specific for anti-atrazine
      monoclonal antibody MoAb K4E7-atrazine immune
      complex (IC)

<400> 7
Cys Pro Ser Ser Arg Trp Phe Asp Leu Cys
 1               5                  10  


<210> 8
<211> 100
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic PCR amplification primer A forward for
      Streptomyces avidinii streptavidin-peptide chimera
      construction

<400> 8
aaaaaaggcc caggcggccg gaggctgtct ggaggcgccg aacattgaag gctgcggtgg 60
cggctctgga ggtggcagtg ccgccgaagc cggcatcacg                       100

<210> 9
<211> 33
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic translation of PCR amplification primer
      A forward for Streptomyces avidinii
      streptavidin-peptide chimera construction

<400> 9
Lys Lys Ala Gln Ala Ala Gly Gly Cys Leu Glu Ala Pro Asn Ile Glu
 1               5                  10                  15      
Gly Cys Gly Gly Gly Ser Gly Gly Gly Ser Ala Ala Glu Ala Gly Ile
            20                  25                  30          
Thr
    

<210> 10
<211> 8
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic (G-3S)-2 spacer

<400> 10
Gly Gly Gly Ser Gly Gly Gly Ser
 1               5              


<210> 11
<211> 112
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic PCR amplification primer B forward for
      Streptomyces avidinii streptavidin-peptide chimera
      construction

<400> 11
tttaaaggcc caggcggccg catccggcag cgcctgctcc ctgtgggaca ccacaggctg 60
gtgtggccct cctcctcctc ctcctggcgc cgccgaagcc ggcatcacgg gc         112

<210> 12
<211> 37
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic translation of PCR amplification primer
      B forward for Streptomyces avidinii
      streptavidin-peptide chimera construction

<400> 12
Leu Lys Ala Gln Ala Ala Ala Ser Gly Ser Ala Cys Ser Leu Trp Asp
 1               5                  10                  15      
Thr Thr Gly Trp Cys Gly Pro Pro Pro Pro Pro Pro Gly Ala Ala Glu
            20                  25                  30          
Ala Gly Ile Thr Gly
        35          


<210> 13
<211> 5
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic ASGSA spacer

<400> 13
Ala Ser Gly Ser Ala
 1               5  


<210> 14
<211> 8
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic GP-6G spacer

<400> 14
Gly Pro Pro Pro Pro Pro Pro Gly
 1               5              


<210> 15
<211> 40
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic PCR amplification primer C reverse for
      Streptomyces avidinii streptavidin-peptide chimera
      construction

<400> 15
gttaaaccgt ctgccgcgtc cggccaggcc ggcctttttt                       40

<210> 16
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic translation of PCR amplification primer
      C reverse for Streptomyces avidinii
      streptavidin-peptide chimera construction

<400> 16
Val Lys Pro Ser Ala Ala Ser Gly Gln Ala Gly Leu Phe
 1               5                  10              


<210> 17
<211> 6
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic His6-tag, histidine tail

<400> 17
His His His His His His
 1               5      


<210> 18
<211> 66
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic OmpA signal sequence and SfiI (1) site
      from pET-OmpA expresssion vector

<400> 18
atgaaaaaga cagctatcgc gattgcagtg gcactggctg gtttcgctac cgtggcccag 60
gcggcc                                                            66

<210> 19
<211> 87
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic SfiI (2) site, His6-tag, HA epitope,
      BamHI, 2 stop codons and XhoI site  from pET-OmpA
      expresssion vector

<400> 19
ggccaggccg gccagcacca tcaccatcac cacggcgcat acccgtacga cgttccggac 60
tacgctagcg gatcctagta gctcgag                                     87

<210> 20
<211> 207
<212> DNA
<213> Enterobacteria phage H-19B

<220> 
<223> verotoxin 1 B subunit (VT1B subunit), Shiga-like
      toxin I subunit B (slt1B)

<400> 20
acgcctgatt gtgtaactgg aaaggtggag tatacaaaat ataatgatga cgataccttt 60
acagttaaag tgggtgataa agaattattt accaacagat ggaatcttca gtctcttctt 120
ctcagtgcgc aaattacggg gatgactgta accattaaaa ctaatgcctg tcataatgga 180
gggggattca gcgaagttat ttttcgt                                     207

<210> 21
<211> 314
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic verotoxin-pA chimera

<400> 21
ggcccaggcg gccggtaccg catccggcag cgcctgctcc ctgtgggaca ccacaggctg 60
gtgtctgcag ggtggcggct ctggaggtgg cagtacgcct gattgtgtaa ctggaaaggt 120
ggagtataca aaatataatg atgacgatac ctttacagtt aaagtgggtg ataaagaatt 180
atttaccaac agatggaatc ttcagtctct tcttctcagt gcgcaaatta cggggatgac 240
tgtaaccatt aaaactaatg cctgtcataa tggaggggga ttcagcgaag ttatttttcg 300
tggccaggcc ggcc                                                   314

<210> 22
<211> 104
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic verotoxin-pA chimera

<400> 22
Ala Gln Ala Ala Gly Thr Ala Ser Gly Ser Ala Cys Ser Leu Trp Asp
 1               5                  10                  15      
Thr Thr Gly Trp Cys Leu Gln Gly Gly Gly Ser Gly Gly Gly Ser Thr
            20                  25                  30          
Pro Asp Cys Val Thr Gly Lys Val Glu Tyr Thr Lys Tyr Asn Asp Asp
        35                  40                  45              
Asp Thr Phe Thr Val Lys Val Gly Asp Lys Glu Leu Phe Thr Asn Arg
    50                  55                  60                  
Trp Asn Leu Gln Ser Leu Leu Leu Ser Ala Gln Ile Thr Gly Met Thr
65                  70                  75                  80  
Val Thr Ile Lys Thr Asn Ala Cys His Asn Gly Gly Gly Phe Ser Glu
                85                  90                  95      
Val Ile Phe Arg Gly Gln Ala Gly
            100                 


<210> 23
<211> 51
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic primer D for verotoxin-pA9 chimera

<400> 23
cgcatccggc agcgcctgtc cgtcgtcgag gtggtttgat ttgtgtctgc a          51

<210> 24
<211> 51
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic primer E for verotoxin-pA9 chimera

<400> 24
gacacaaatc aaaccacctc gacgacggac aggcgctgcc ggatgcggta c          51

<210> 25
<211> 534
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic streptavidin-pA nanopeptamer subunit

<400> 25
gcatccggca gcgcctgctc cctgtgggac accacaggct ggtgtggccc tcctcctcct 60
cctcctggcg ccgccgaagc cggcatcacg ggcacttggt acaaccagct gggtagcacc 120
tttattgtca ccgcgggcgc cgatggggca ctgacaggta cctatgagag cgctgtgggc 180
aacgcagaga gccgctatgt gctgaccggg cgctatgatt ccgccccggc caccgatggt 240
tctggtacgg ccctgggttg gacggtggct tggaagaata actatcgtaa cgcgcattcc 300
gccactacct ggtccggtca atatgtgggc ggggccgaag cacgtattaa tacccagtgg 360
ctgttaacct ctggtacgac cgaagccaac gcctggaaga gcaccctggt cggccatgat 420
acgtttacca aagttaaacc gtctgccgcg tccggccagg ccggccagca ccatcaccat 480
caccacggcg catacccgta cgacgttccg gactacgcta gcggatccta gtag       534

<210> 26
<211> 176
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic streptavidin-pA nanopeptamer subunit

<400> 26
Ala Ser Gly Ser Ala Cys Ser Leu Trp Asp Thr Thr Gly Trp Cys Gly
 1               5                  10                  15      
Pro Pro Pro Pro Pro Pro Gly Ala Ala Glu Ala Gly Ile Thr Gly Thr
            20                  25                  30          
Trp Tyr Asn Gln Leu Gly Ser Thr Phe Ile Val Thr Ala Gly Ala Asp
        35                  40                  45              
Gly Ala Leu Thr Gly Thr Tyr Glu Ser Ala Val Gly Asn Ala Glu Ser
    50                  55                  60                  
Arg Tyr Val Leu Thr Gly Arg Tyr Asp Ser Ala Pro Ala Thr Asp Gly
65                  70                  75                  80  
Ser Gly Thr Ala Leu Gly Trp Thr Val Ala Trp Lys Asn Asn Tyr Arg
                85                  90                  95      
Asn Ala His Ser Ala Thr Thr Trp Ser Gly Gln Tyr Val Gly Gly Ala
            100                 105                 110         
Glu Ala Arg Ile Asn Thr Gln Trp Leu Leu Thr Ser Gly Thr Thr Glu
        115                 120                 125             
Ala Asn Ala Trp Lys Ser Thr Leu Val Gly His Asp Thr Phe Thr Lys
    130                 135                 140                 
Val Lys Pro Ser Ala Ala Ser Gly Gln Ala Gly Gln His His His His
145                 150                 155                 160 
His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Gly Ser
                165                 170                 175     


<210> 27
<211> 525
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic streptavidin-pICX11 nanopeptamer subunit

<400> 27
ggaggctgtc tggaggcgcc gaacattgaa ggctgcggtg gcggctctgg aggtggcagt 60
gccgccgaag ccggcatcac gggcacttgg tacaaccagc tgggtagcac ctttattgtc 120
accgcgggcg ccgatggggc actgacaggt acctatgaga gcgctgtggg caacgcagag 180
agccgctatg tgctgaccgg gcgctatgat tccgccccgg ccaccgatgg ttctggtacg 240
gccctgggtt ggacggtggc ttggaagaat aactatcgta acgcgcattc cgccactacc 300
tggtccggtc aatatgtggg cggggccgaa gcacgtatta atacccagtg gctgttaacc 360
tctggtacga ccgaagccaa cgcctggaag agcaccctgg tcggccatga tacgtttacc 420
aaagttaaac cgtctgccgc gtccggccag gccggccagc accatcacca tcaccacggc 480
gcatacccgt acgacgttcc ggactacgct agcggatcct agtag                 525

<210> 28
<211> 173
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic streptavidin-pICX11 nanopeptamer subunit

<400> 28
Gly Gly Cys Leu Glu Ala Pro Asn Ile Glu Gly Cys Gly Gly Gly Ser
 1               5                  10                  15      
Gly Gly Gly Ser Ala Ala Glu Ala Gly Ile Thr Gly Thr Trp Tyr Asn
            20                  25                  30          
Gln Leu Gly Ser Thr Phe Ile Val Thr Ala Gly Ala Asp Gly Ala Leu
        35                  40                  45              
Thr Gly Thr Tyr Glu Ser Ala Val Gly Asn Ala Glu Ser Arg Tyr Val
    50                  55                  60                  
Leu Thr Gly Arg Tyr Asp Ser Ala Pro Ala Thr Asp Gly Ser Gly Thr
65                  70                  75                  80  
Ala Leu Gly Trp Thr Val Ala Trp Lys Asn Asn Tyr Arg Asn Ala His
                85                  90                  95      
Ser Ala Thr Thr Trp Ser Gly Gln Tyr Val Gly Gly Ala Glu Ala Arg
            100                 105                 110         
Ile Asn Thr Gln Trp Leu Leu Thr Ser Gly Thr Thr Glu Ala Asn Ala
        115                 120                 125             
Trp Lys Ser Thr Leu Val Gly His Asp Thr Phe Thr Lys Val Lys Pro
    130                 135                 140                 
Ser Ala Ala Ser Gly Gln Ala Gly Gln His His His His His His Gly
145                 150                 155                 160 
Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Gly Ser
                165                 170             


<210> 29
<211> 369
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic verotoxin-pA nanopeptamer subunit

<400> 29
ggtaccgcat ccggcagcgc ctgctccctg tgggacacca caggctggtg tctgcagggt 60
ggcggctctg gaggtggcag tacgcctgat tgtgtaactg gaaaggtgga gtatacaaaa 120
tataatgatg acgatacctt tacagttaaa gtgggtgata aagaattatt taccaacaga 180
tggaatcttc agtctcttct tctcagtgcg caaattacgg ggatgactgt aaccattaaa 240
actaatgcct gtcataatgg agggggattc agcgaagtta tttttcgtgg ccaggccggc 300
cagcaccatc accatcacca cggcgcatac ccgtacgacg ttccggacta cgctagcgga 360
tcctagtag                                                         369

<210> 30
<211> 121
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic verotoxin-pA nanopeptamer subunit

<400> 30
Gly Thr Ala Ser Gly Ser Ala Cys Ser Leu Trp Asp Thr Thr Gly Trp
 1               5                  10                  15      
Cys Leu Gln Gly Gly Gly Ser Gly Gly Gly Ser Thr Pro Asp Cys Val
            20                  25                  30          
Thr Gly Lys Val Glu Tyr Thr Lys Tyr Asn Asp Asp Asp Thr Phe Thr
        35                  40                  45              
Val Lys Val Gly Asp Lys Glu Leu Phe Thr Asn Arg Trp Asn Leu Gln
    50                  55                  60                  
Ser Leu Leu Leu Ser Ala Gln Ile Thr Gly Met Thr Val Thr Ile Lys
65                  70                  75                  80  
Thr Asn Ala Cys His Asn Gly Gly Gly Phe Ser Glu Val Ile Phe Arg
                85                  90                  95      
Gly Gln Ala Gly Gln His His His His His His Gly Ala Tyr Pro Tyr
            100                 105                 110         
Asp Val Pro Asp Tyr Ala Ser Gly Ser
        115                 120     


<210> 31
<211> 369
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetic verotoxin-pA9 nanopeptamer subunit

<400> 31
ggtaccgcat ccggcagcgc ctgtccgtcg tcgaggtggt ttgatttgtg tctgcagggt 60
ggcggctctg gaggtggcag tacgcctgat tgtgtaactg gaaaggtgga gtatacaaaa 120
tataatgatg acgatacctt tacagttaaa gtgggtgata aagaattatt taccaacaga 180
tggaatcttc agtctcttct tctcagtgcg caaattacgg ggatgactgt aaccattaaa 240
actaatgcct gtcataatgg agggggattc agcgaagtta tttttcgtgg ccaggccggc 300
cagcaccatc accatcacca cggcgcatac ccgtacgacg ttccggacta cgctagcgga 360
tcctagtag                                                         369

<210> 32
<211> 121
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetic verotoxin-pA9 nanopeptamer subunit

<400> 32
Gly Thr Ala Ser Gly Ser Ala Cys Pro Ser Ser Arg Trp Phe Asp Leu
 1               5                  10                  15      
Cys Leu Gln Gly Gly Gly Ser Gly Gly Gly Ser Thr Pro Asp Cys Val
            20                  25                  30          
Thr Gly Lys Val Glu Tyr Thr Lys Tyr Asn Asp Asp Asp Thr Phe Thr
        35                  40                  45              
Val Lys Val Gly Asp Lys Glu Leu Phe Thr Asn Arg Trp Asn Leu Gln
    50                  55                  60                  
Ser Leu Leu Leu Ser Ala Gln Ile Thr Gly Met Thr Val Thr Ile Lys
65                  70                  75                  80  
Thr Asn Ala Cys His Asn Gly Gly Gly Phe Ser Glu Val Ile Phe Arg
                85                  90                  95      
Gly Gln Ala Gly Gln His His His His His His Gly Ala Tyr Pro Tyr
            100                 105                 110         
Asp Val Pro Asp Tyr Ala Ser Gly Ser
        115                 120     

