
                                SEQUENCE LISTING

<110> Pioneer Hi-Bred International, Inc.
      
<120> Tetracycline Repressor and Uses Thereof
  

<130> 2011-PCT

<150> 60/866,876          
<151> 2006-11-22  

<160> 16

<170> FastSEQ for Windows Version 4.0

<210> 1
<211> 624
<212> DNA
<213> E. coli

<400> 1
atgtctagat tagataaaag taaagtgatt aacagcgcat tagagctgct taatgaggtc 60
ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga gcagcctaca 120
ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat tgagatgtta 180
gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga ttttttacgt 240
aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc aaaagtacat 300
ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt agccttttta 360
tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgctgt ggggcatttt 420
actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga aagggaaaca 480
cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt tgatcaccaa 540
ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt agaaaaacaa 600
cttaaatgtg aaagtgggtc ttaa                                        624

<210> 2
<211> 207
<212> PRT
<213> E. coli

<400> 2
Met Ser Arg Leu Asp Lys Ser Lys Val Ile Asn Ser Ala Leu Glu Leu
 1               5                  10                  15      
Leu Asn Glu Val Gly Ile Glu Gly Leu Thr Thr Arg Lys Leu Ala Gln
            20                  25                  30          
Lys Leu Gly Val Glu Gln Pro Thr Leu Tyr Trp His Val Lys Asn Lys
        35                  40                  45              
Arg Ala Leu Leu Asp Ala Leu Ala Ile Glu Met Leu Asp Arg His His
    50                  55                  60                  
Thr His Phe Cys Pro Leu Glu Gly Glu Ser Trp Gln Asp Phe Leu Arg
65                  70                  75                  80  
Asn Asn Ala Lys Ser Phe Arg Cys Ala Leu Leu Ser His Arg Asp Gly
                85                  90                  95      
Ala Lys Val His Leu Gly Thr Arg Pro Thr Glu Lys Gln Tyr Glu Thr
            100                 105                 110         
Leu Glu Asn Gln Leu Ala Phe Leu Cys Gln Gln Gly Phe Ser Leu Glu
        115                 120                 125             
Asn Ala Leu Tyr Ala Leu Ser Ala Val Gly His Phe Thr Leu Gly Cys
    130                 135                 140                 
Val Leu Glu Asp Gln Glu His Gln Val Ala Lys Glu Glu Arg Glu Thr
145                 150                 155                 160 
Pro Thr Thr Asp Ser Met Pro Pro Leu Leu Arg Gln Ala Ile Glu Leu
                165                 170                 175     
Phe Asp His Gln Gly Ala Glu Pro Ala Phe Leu Phe Gly Leu Glu Leu
            180                 185                 190         
Ile Ile Cys Gly Leu Glu Lys Gln Leu Lys Cys Glu Ser Gly Ser
        195                 200                 205         


<210> 3
<211> 624
<212> DNA
<213> Artificial Sequence

<220> 
<223> Codon modified TetRMOD1

<400> 3
atgtctagac tcgacaagag caaggtgatc aacagcgcac tcgagctgct gaacgaggtc 60
ggaatcgaag gcctcacaac ccgtaaactc gcccagaagc tcggtgtaga gcagcctaca 120
ttgtattggc atgtcaagaa caagcgggct ttgctcgacg ccctcgccat tgagatgctc 180
gataggcacc atacccactt ctgccctttg gaaggggaaa gctggcaaga cttcctgcgc 240
aacaacgcta agagcttcag atgtgctttg ctcagtcatc gcgatggagc caaggtccac 300
ttgggtacac ggcctacaga gaagcagtat gaaactctcg agaaccagct cgccttcctg 360
tgccaacaag gtttctccct tgagaatgcc ctctacgcac tcagcgctgt ggggcacttc 420
actctgggtt gcgtattgga agatcaagag catcaagtcg ctaaggagga gagggaaaca 480
cctactactg atagtatgcc gccactgctc cgacaagcta tcgagctctt cgatcaccaa 540
ggtgcagagc cagccttcct gttcggcctt gaattgatca tatgcggatt ggagaagcag 600
ctgaagtgtg aaagtgggtc ttaa                                        624

<210> 4
<211> 207
<212> PRT
<213> Artificial Sequence

<220> 
<223> Tetracycline repressor encoded by codon modified
      TetRMOD1

<400> 4
Met Ser Arg Leu Asp Lys Ser Lys Val Ile Asn Ser Ala Leu Glu Leu
 1               5                  10                  15      
Leu Asn Glu Val Gly Ile Glu Gly Leu Thr Thr Arg Lys Leu Ala Gln
            20                  25                  30          
Lys Leu Gly Val Glu Gln Pro Thr Leu Tyr Trp His Val Lys Asn Lys
        35                  40                  45              
Arg Ala Leu Leu Asp Ala Leu Ala Ile Glu Met Leu Asp Arg His His
    50                  55                  60                  
Thr His Phe Cys Pro Leu Glu Gly Glu Ser Trp Gln Asp Phe Leu Arg
65                  70                  75                  80  
Asn Asn Ala Lys Ser Phe Arg Cys Ala Leu Leu Ser His Arg Asp Gly
                85                  90                  95      
Ala Lys Val His Leu Gly Thr Arg Pro Thr Glu Lys Gln Tyr Glu Thr
            100                 105                 110         
Leu Glu Asn Gln Leu Ala Phe Leu Cys Gln Gln Gly Phe Ser Leu Glu
        115                 120                 125             
Asn Ala Leu Tyr Ala Leu Ser Ala Val Gly His Phe Thr Leu Gly Cys
    130                 135                 140                 
Val Leu Glu Asp Gln Glu His Gln Val Ala Lys Glu Glu Arg Glu Thr
145                 150                 155                 160 
Pro Thr Thr Asp Ser Met Pro Pro Leu Leu Arg Gln Ala Ile Glu Leu
                165                 170                 175     
Phe Asp His Gln Gly Ala Glu Pro Ala Phe Leu Phe Gly Leu Glu Leu
            180                 185                 190         
Ile Ile Cys Gly Leu Glu Lys Gln Leu Lys Cys Glu Ser Gly Ser
        195                 200                 205         


<210> 5
<211> 624
<212> DNA
<213> Artificial Sequence

<220> 
<223> Codon modified TetRMOD3

<400> 5
atggccagac tcgacaagag caaggtgatc aacagcgcac tcgagctgct gaacgaggtc 60
ggaatcgaag gcctcacaac ccgtaaactc gcccagaagc tcggggtaga gcagcctaca 120
ttgtattggc atgtcaagaa caagcgggct ttgctcgacg ccctcgccat tgagatgctc 180
gataggcacc atacccactt ctgccctttg gaaggggaaa gctggcaaga cttcctgcgc 240
aacaacgcta agagcttcag atgtgctttg ctcagtcatc gcgatggagc caaggtccac 300
ttgggtacac ggcctacgga gaagcagtat gaaactctcg agaaccagct cgccttcctg 360
tgccaacaag gtttctccct tgagaatgcc ctctacgcac tctccgctgt ggggcacttc 420
actctgggtt gcgtattgga agatcaagag catcaagtcg ctaaggagga gagggaaaca 480
cctactactg atagtatgcc gccactgctc cgacaagcta tcgagctctt cgatcaccaa 540
ggtgcagagc cagccttcct gttcggcctt gaattgatca tatgcggatt ggagaagcag 600
ctgaagtgtg aaagtgggtc ttaa                                        624

<210> 6
<211> 207
<212> PRT
<213> Artificial Sequence

<220> 
<223> Tetracycline repressor encoded by codon modified
      TetRMOD3

<400> 6
Met Ala Arg Leu Asp Lys Ser Lys Val Ile Asn Ser Ala Leu Glu Leu
 1               5                  10                  15      
Leu Asn Glu Val Gly Ile Glu Gly Leu Thr Thr Arg Lys Leu Ala Gln
            20                  25                  30          
Lys Leu Gly Val Glu Gln Pro Thr Leu Tyr Trp His Val Lys Asn Lys
        35                  40                  45              
Arg Ala Leu Leu Asp Ala Leu Ala Ile Glu Met Leu Asp Arg His His
    50                  55                  60                  
Thr His Phe Cys Pro Leu Glu Gly Glu Ser Trp Gln Asp Phe Leu Arg
65                  70                  75                  80  
Asn Asn Ala Lys Ser Phe Arg Cys Ala Leu Leu Ser His Arg Asp Gly
                85                  90                  95      
Ala Lys Val His Leu Gly Thr Arg Pro Thr Glu Lys Gln Tyr Glu Thr
            100                 105                 110         
Leu Glu Asn Gln Leu Ala Phe Leu Cys Gln Gln Gly Phe Ser Leu Glu
        115                 120                 125             
Asn Ala Leu Tyr Ala Leu Ser Ala Val Gly His Phe Thr Leu Gly Cys
    130                 135                 140                 
Val Leu Glu Asp Gln Glu His Gln Val Ala Lys Glu Glu Arg Glu Thr
145                 150                 155                 160 
Pro Thr Thr Asp Ser Met Pro Pro Leu Leu Arg Gln Ala Ile Glu Leu
                165                 170                 175     
Phe Asp His Gln Gly Ala Glu Pro Ala Phe Leu Phe Gly Leu Glu Leu
            180                 185                 190         
Ile Ile Cys Gly Leu Glu Lys Gln Leu Lys Cys Glu Ser Gly Ser
        195                 200                 205         


<210> 7
<211> 447
<212> DNA
<213> Artificial Sequence

<220> 
<223> 35S Triple-Op promoter

<400> 7
gaaaatcttc gtcaacatgg tggagcacga cacgcttgtc tactccaaaa atatcaaaga 60
tacagtctca gaagaccaaa gggcaattga gacttttcaa caaagggtaa tatccggaaa 120
cctcctcgga ttccattgcc cagctatctg tcactttatt gtgaagatag tggaaaagga 180
aggtggctcc tacaaatgcc atcattgcga taaaggaaag gccatcgttg aagatgcctc 240
tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga 300
cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat atctccactg acgtaaggga 360
tgacgcacaa tcccactaga ctctatcagt gatagagtgt atataagact ctatcagtga 420
tagagtgaac tctatcagtg atagagt                                     447

<210> 8
<211> 33
<212> DNA
<213> Artificial Sequence

<220> 
<223> PCR primer - Forward primer PHN107613 to omega 5'
      UTR

<400> 8
tacaacaatt accaacaaca acaaacaaca aac                              33

<210> 9
<211> 21
<212> DNA
<213> Artificial Sequence

<220> 
<223> PCR primer - reverse primer PHN107611 to ODP2

<400> 9
agaaggagat gccgccgagg a                                           21

<210> 10
<211> 19
<212> DNA
<213> Artificial Sequence

<220> 
<223> Tetracycline Operator used in 35S Triple-Op

<400> 10
actctatcag tgatagagt                                              19

<210> 11
<211> 624
<212> DNA
<213> Artificial Sequence

<220> 
<223> Codon modified TetRMOD4

<400> 11
atggccagac tcgacaagag caaggtgatc aacagcgcac tggagctgct gaacgaggtc 60
ggaatcgaag gcctcacaac ccgtaaactc gcccagaagc tcggggtaga gcagcctaca 120
ttgtattggc acgtcaagaa caagcgggct ttgctagacg ccctcgccat tgagatgctc 180
gataggcacc atacccactt ctgccctttg gaaggggaaa gctggcaaga cttcttgagg 240
aacaacgcta agagcttcag atgtgctttg ctcagtcacc gtgatggagc caaggtccac 300
ctaggtacac ggcctacgga gaagcagtat gaaactctcg agaaccagct cgccttcctg 360
tgccaacaag gtttctccct tgagaatgcc ctctacgcac tctccgctgt agggcacttc 420
actctgggtt gcgtattgga agatcaagag caccaagtcg ctaaggagga gagggaaaca 480
cctactactg atagtatgcc gccactgctc cgacaagcta tcgagctctt cgatcaccaa 540
ggtgcagagc cagccttcct gttcggcctt gaattgatca tatgcggatt ggagaagcag 600
ctgaagtgtg aaagtgggtc ttaa                                        624

<210> 12
<211> 207
<212> PRT
<213> Artificial Sequence

<220> 
<223> Tetracycline repressor encoded by codon modified
      TetRMOD4

<400> 12
Met Ala Arg Leu Asp Lys Ser Lys Val Ile Asn Ser Ala Leu Glu Leu
 1               5                  10                  15      
Leu Asn Glu Val Gly Ile Glu Gly Leu Thr Thr Arg Lys Leu Ala Gln
            20                  25                  30          
Lys Leu Gly Val Glu Gln Pro Thr Leu Tyr Trp His Val Lys Asn Lys
        35                  40                  45              
Arg Ala Leu Leu Asp Ala Leu Ala Ile Glu Met Leu Asp Arg His His
    50                  55                  60                  
Thr His Phe Cys Pro Leu Glu Gly Glu Ser Trp Gln Asp Phe Leu Arg
65                  70                  75                  80  
Asn Asn Ala Lys Ser Phe Arg Cys Ala Leu Leu Ser His Arg Asp Gly
                85                  90                  95      
Ala Lys Val His Leu Gly Thr Arg Pro Thr Glu Lys Gln Tyr Glu Thr
            100                 105                 110         
Leu Glu Asn Gln Leu Ala Phe Leu Cys Gln Gln Gly Phe Ser Leu Glu
        115                 120                 125             
Asn Ala Leu Tyr Ala Leu Ser Ala Val Gly His Phe Thr Leu Gly Cys
    130                 135                 140                 
Val Leu Glu Asp Gln Glu His Gln Val Ala Lys Glu Glu Arg Glu Thr
145                 150                 155                 160 
Pro Thr Thr Asp Ser Met Pro Pro Leu Leu Arg Gln Ala Ile Glu Leu
                165                 170                 175     
Phe Asp His Gln Gly Ala Glu Pro Ala Phe Leu Phe Gly Leu Glu Leu
            180                 185                 190         
Ile Ile Cys Gly Leu Glu Lys Gln Leu Lys Cys Glu Ser Gly Ser
        195                 200                 205         


<210> 13
<211> 624
<212> DNA
<213> Artificial Sequence

<220> 
<223> Codon modified TetRMOD2

<400> 13
atgtctagac tcgacaagag caaggtgatc aacagcgcac tcgagctgct gaacgaggtc 60
ggaatcgaag gcctcacaac ccgtaaactc gcccagaagc tcggtgtaga gcagcctaca 120
ttgtattggc acgtcaagaa caagcgggct ttgctagacg ccctcgccat tgagatgctc 180
gataggcacc atacccactt ctgccctttg gaaggggaaa gctggcaaga cttcttgagg 240
aacaacgcta agagcttcag atgtgctttg ctcagtcatc gcgatggagc caaggtccac 300
ttgggtacac ggcctacaga gaagcagtat gaaactctcg agaaccagct cgccttcctg 360
tgccaacaag gtttctccct tgagaatgcc ctctacgcac tcagcgctgt agggcacttc 420
actctgggtt gcgtattgga agatcaagag catcaagtcg ctaaggagga gagggaaaca 480
cctactactg atagtatgcc gccactgctc cgacaagcta tcgagctctt cgatcaccaa 540
ggtgcagagc cagccttcct gttcggcctt gaattgatca tatgcggatt ggagaagcag 600
ctgaagtgtg aaagtgggtc ttaa                                        624

<210> 14
<211> 207
<212> PRT
<213> Artificial Sequence

<220> 
<223> Tetracycline repressor encoded by codon modified
      TetRMOD2

<400> 14
Met Ser Arg Leu Asp Lys Ser Lys Val Ile Asn Ser Ala Leu Glu Leu
 1               5                  10                  15      
Leu Asn Glu Val Gly Ile Glu Gly Leu Thr Thr Arg Lys Leu Ala Gln
            20                  25                  30          
Lys Leu Gly Val Glu Gln Pro Thr Leu Tyr Trp His Val Lys Asn Lys
        35                  40                  45              
Arg Ala Leu Leu Asp Ala Leu Ala Ile Glu Met Leu Asp Arg His His
    50                  55                  60                  
Thr His Phe Cys Pro Leu Glu Gly Glu Ser Trp Gln Asp Phe Leu Arg
65                  70                  75                  80  
Asn Asn Ala Lys Ser Phe Arg Cys Ala Leu Leu Ser His Arg Asp Gly
                85                  90                  95      
Ala Lys Val His Leu Gly Thr Arg Pro Thr Glu Lys Gln Tyr Glu Thr
            100                 105                 110         
Leu Glu Asn Gln Leu Ala Phe Leu Cys Gln Gln Gly Phe Ser Leu Glu
        115                 120                 125             
Asn Ala Leu Tyr Ala Leu Ser Ala Val Gly His Phe Thr Leu Gly Cys
    130                 135                 140                 
Val Leu Glu Asp Gln Glu His Gln Val Ala Lys Glu Glu Arg Glu Thr
145                 150                 155                 160 
Pro Thr Thr Asp Ser Met Pro Pro Leu Leu Arg Gln Ala Ile Glu Leu
                165                 170                 175     
Phe Asp His Gln Gly Ala Glu Pro Ala Phe Leu Phe Gly Leu Glu Leu
            180                 185                 190         
Ile Ile Cys Gly Leu Glu Lys Gln Leu Lys Cys Glu Ser Gly Ser
        195                 200                 205         


<210> 15
<211> 19
<212> DNA
<213> E. coli

<220> 
<221> protein_bind   
<222> (1)...(19)
<223> Tetracycline Operator O1

<400> 15
actctatcat tgatagagt                                              19

<210> 16
<211> 19
<212> DNA
<213> E. coli

<220> 
<221> protein_bind   
<222> (1)...(19)
<223> Tetracycline Operator O2

<400> 16
tccctatcag tgatagaga                                              19
