IGEM:IMPERIAL/2009/Primers

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m (Primers to be used for PCR)
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Forward: <b>GCTCTAG</b>ATGAGTCGTTTAGTCGTAG &nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 52.4C(without) and 63.2(with)<br>
Forward: <b>GCTCTAG</b>ATGAGTCGTTTAGTCGTAG &nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 52.4C(without) and 63.2(with)<br>
Backward: <b>GGACTAGTA</b>CTACGCAAGCTTTGGAAAG &nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 54.5C(without) and 65.1(with)<br>
Backward: <b>GGACTAGTA</b>CTACGCAAGCTTTGGAAAG &nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 54.5C(without) and 65.1(with)<br>
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Strain to use:BL21(DE3)
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<b>PAH Sequence:</b>
<b>PAH Sequence:</b>

Revision as of 09:52, 10 August 2009

Primers to be used for PCR

Considerations for primer design http://www.premierbiosoft.com/tech_notes/PCR_Primer_Design.html

Use this website to calculate temperatures and modify primers and this one for reverse complement.


RcsB Sequence:

     ATGAACAATATGAACGTAATTATTGCCGATGACCATCCGATAGTCTTGTTCGGTATTCGCAAATCACTTG
     AGCAAATTGAGTGGGTGAATGTTGTCGGCGAATTTGAAGACTCTACAGCACTGATCAACAACCTGCCGAA
     ACTGGATGCGCATGTGTTGATTACCGATCTCTCCATGCCTGGCGATAAGTACGGCGATGGCATTACCTTA
     ATCAAGTACATCAAGCGCCATTTCCCAAGCCTGTCGATCATTGTTCTGACTATGAACAACAACCCGGCGA
     TTCTTAGTGCGGTATTGGATCTGGATATCGAAGGGATCGTGCTGAAACAAGGTGCACCGACCGATCTGCC
     GAAAGCTCTCGCCGCGCTGCAGAAAGGGAAGAAATTTACCCCGGAAAGCGTTTCTCGCCTGTTGGAAAAA
     ATCAGTGCTGGTGGTTACGGTGACAAGCGTCTCTCGCCAAAAGAGAGTGAAGTTCTGCGCCTGTTTGCGG
     AAGGCTTCCTGGTGACCGAGATCGCTAAAAAGCTGAACCGCAGTATTAAAACCATCAGTAGCCAGAAGAA
     ATCTGCGATGATGAAGCTGGGTGTCGAGAACGATATCGCCCTGCTGAATTATCTCTCTTCAGTGACCTTA
     AGTCCGGCAGATAAAGACTAA

Forward: GCTCTAGATGAACAATATGAACGTAATTATTG       53.1C(without) and 61.8(with)
Backward: GGACTAGTATTAGTCTTTATCTGCCGG       51.4C(without) and 63.4(with)

Strains to use:DH10B/TOP10
OtsB Sequence:

      ATGACAGAACCGTTAACCGAAACCCCTGAACTATCCGCGAAATATGCCTGGTTTTTTGATCTTGATG
      GAACGCTGGCGGAAATCAAACCGCATCCCGATCAGGTCGTCGTGCCTGACAATATTCTGCAAGGACT
      ACAGCTACTGGCAACCGCAAGTGATGGTGCATTGGCATTGATATCAGGGCGCTCAATGGTGGAGCTT
      GACGCACTGGCAAAACCTTATCGCTTCCCGTTAGCGGGCGTGCATGGGGCGGAGCGCCGTGACATCA
      ATGGTAAAACACATATCGTTCATCTGCCGGATGCGATTGCGCGTGATATTAGCGTGCAACTGCATAC
      AGTCATCGCTCAGTATCCCGGCGCGGAGCTGGAGGCGAAAGGGATGGCTTTTGCGCTGCATTATCGT
      CAGGCTCCGCAGCATGAAGACGCATTAATGACATTAGCGCAACGTATTACTCAGATCTGGCCACAAA
      TGGCGTTACAGCAGGGAAAGTGTGTTGTCGAGATCAAACCGAGAGGTACCAGTAAAGGTGAGGCAAT
      TGCAGCTTTTATGCAGGAAGCTCCCTTTATCGGGCGAACGCCCGTATTTCTGGGCGATGATTTAACC
      GATGAATCTGGCTTCGCAGTCGTTAACCGACTGGGCGGAATGTCAGTAAAAATTGGCACAGGTGCAA
      CTCAGGCATCATGGCGACTGGCGGGTGTGCCGGATGTCTGGAGCTGGCTTGAAATGATAACCACCGC
      ATTACAACAAAAAAGAGAAAATAACAGGAGTGATGACTATGAGTCGTTTAGTCGTAGTATCTAA


Forward: GCTCTAGATGACAGAACCGTTAACCG       54.5C (without) & 64.8C (with)
Backward: GGACTAGTATTAGATACTACGACTAAACGAC       54.7C (without) & 64.2C (with)


Strains to use:BL21(DE3)
B3023 Sequence:

     ATGACAAACCTGACACTGGATGTAAACATTATCGATTTCCCATCAATACCTGTGGCGATGTTGCCG
     CACCGCTGTAGCCCTGAATTGCTCAACTACAGCGTGGCGAAATTTATCATGTGGCGTAAAGAGACG
     GGGCTTTCTCCTGTTAACCAAAGCCAGACTTTTGGCGTCGCCTGGGACGACCCTGCCACCACCGCA
     CCGGAAGCGTTTCGCTTTGATATCTGCGGCAGCGTTAGCGAACCGATTCCCGATAATCGTTATGGT
     GTGAGCAATGGTGAACTTACCGGTGGACGTTATGCCGTGGCCCGCCACGTTGGCGAGCTGGACGAT
     ATTTCACACACGGTATGGGGCATCATTCGCCACTGGCTGCCTGCAAGCGGCGAGAAAATGCGTAAA
     GCACCGATTCTGTTTCACTACACCAATCTTGCCGAAGGGGTGACAGAGCAGCGACTGGAAACGGAT
     GTTTATGTGCCGTTGGCGTGA


Forward: GCTCTAGATGACAAACCTGACACTG       51.4C(without) and 63.0(with)
Backward: GGACTAGTATCACGCCAACGGCAC       53.3C(without) and 66.1C(with)

Strains to use:BL21(DE3)

WaaL Sequence:

      ATGCTAACATCCTTTAAACTTCATTCATTGAAACCTTACACTCTGAAATCATCAATGATTTTAGAG
      ATAATAACTTATATATTATGTTTTTTTTCAATGATAATTGCATTCGTCGATAATACTTTCAGCATA
      AAAATATATAATATCACTGCTATAGTTTGCTTATTGTCACTAATTTTACGTGGCAGACAAGAAAATT
      ATAATATAAAAAACCTTATTCTTCCCCTTTCTATATTTTTAATAGGCTTGCTTGATTTAATTTGGTA
      TTCTGCGTTTAAAGTAGATAATTCGCCATTTCGTGCTACTTACCATAGTTATTTAAATACTGCCAAA
      ATATTTATATTTGGTTCTTTTATTGTTTTCTTGACACTAACTAGCCAGCTAAAATCAAAAAAAGAGA
      GTGTATTATACACTTTGTATTCTCTGTCATTTCTAATTGCTGGATATGCAATGTATATTAATAGCAT
      TCATGAAAATGACCGCATTTCTTTTGGTGTAGGAACGGCAACAGGAGCAGCATATTCAACAATGCTA
      ATAGGGATAGTTAGTGGCGTTGCGATTCTTTATACTAAGAAAAATCATCCTTTTTTATTTTTATTAA
      ATAGTTGCGCGGTACTTTATGTTCTGGCGCTAACACAAACCAGAGCAACCCTACTCCTGTTCCCTAT
      AATTTGTGTTGCTGCATTAATAGCTTATTATAATAAATCACCCAAGAAATTCACTTCCTCTATTGTT
      CTACTAATTGCTATATTAGCTAGCATTGTTATTATATTTAATAAACCAATACAGAATCGCTATAATG
      AAGCATTAAATGACTTAAACAGTTATACCAATGCTAATAGTGTTACTTCCCTAGGTGCAAGACTGGC
      AATGTACGAAATTGGTTTAAATATATTCATAAAGTCACCTTTTTCATTTAGATCAGCAGAGTCACGC
      GCTGAAAGTATGAATTTGTTAGTTGCAGAACACAATAGGCTAAGAGGGGCATTGGAGTTTTCTAACG
      TACATCTACATAATGAGATAATTGAAGCAGGGTCACTGAAAGGTCTGATGGGAATTTTTTCCACACT
      TTTCCTCTATTTTTCACTATTTTATATAGCATATAAAAAACGAGCTTTGGGTTTGTTGATATTAACG
      CTTGGCATTGTGGGGATTGGACTCAGTGATGTGATCATATGGGCACGCAGCATTCCAATTATCATTA
      TATCCGCTATAGTCCTCTTACTCGTCATTAATAATCGTAACAATACAATTAATTAA


Forward: GCTCTAGATGCTAACATCCTTTAAACTTC       52.8C(without) and 62.4(with)
Backward: GGACTAGTATTAATTAATTGTATTGTTACGATTATTAATG       54.9C(without) and 62.3(with)

Strains to use:DH10B/TOP10
OtsA Sequence:

      ATGAGTCGTTTAGTCGTAGTATCTAACCGGATTGCACCACCAGACGAGCACGCCGCCAGTGCCGGTGGCCTTGC
      CGTTGGCATACTGGGGGCACTGAAAGCCGCAGGCGGACTGTGGTTTGGCTGGAGTGGTGAAACAGGGAATGAGG
      ATCAGCCGCTAAAAAAGGTGAAAAAAGGTAACATTACGTGGGCCTCTTTTAACCTCAGCGAACAGGACCTTGAC
      GAATACTACAACCAATTCTCCAATGCCGTTCTCTGGCCCGCTTTTCATTATCGGCTCGATCTGGTGCAATTTCA
      GCGTCCTGCCTGGGACGGCTATCTACGCGTAAATGCGTTGCTGGCAGATAAATTACTGCCGCTGTTGCAAGACG
      ATGACATTATCTGGATCCACGATTATCACCTGTTGCCATTTGCGCATGAATTACGCAAACGGGGAGTGAATAAT
      CGCATTGGTTTCTTTCTGCATATTCCTTTCCCGACACCGGAAATCTTCAACGCGCTGCCGACATATGACACCTT
      GCTTGAACAGCTTTGTGATTATGATTTGCTGGGTTTCCAGACAGAAAACGATCGTCTGGCGTTCCTGGATTGTC
      TTTCTAACCTGACCCGCGTCACGACACGTAGCGCAAAAAGCCATACAGCCTGGGGCAAAGCATTTCGAACAGAA
      GTCTACCCGATCGGCATTGAACCGAAAGAAATAGCCAAACAGGCTGCCGGGCCACTGCCGCCAAAACTGGCGCA
      ACTTAAAGCGGAACTGAAAAACGTACAAAATATCTTTTCTGTCGAACGGCTGGATTATTCCAAAGGTTTGCCAG
      AGCGTTTTCTCGCCTATGAAGCGTTGCTGGAAAAATATCCGCAGCATCATGGTAAAATTCGTTATACCCAGATT
      GCACCAACGTCGCGTGGTGATGTGCAAGCCTATCAGGATATTCGTCATCAGCTCGAAAATGAAGCTGGACGAAT
      TAATGGTAAATACGGGCAATTAGGCTGGACGCCGCTTTATTATTTGAATCAGCATTTTGACCGTAAATTACTGA
      TGAAAATATTCCGCTACTCTGACGTGGGCTTAGTGACGCCACTGCGTGACGGGATGAACCTGGTAGCAAAAGAG
      TATGTTGCTGCTCAGGACCCAGCCAATCCGGGCGTTCTTGTTCTTTCGCAATTTGCGGGAGCGGCAAACGAGTT
      AACGTCGGCGTTAATTGTTAACCCCTACGATCGTGACGAAGTTGCAGCTGCGCTGGATCGTGCATTGACTATGT
      CGCTGGCGGAACGTATTTCCCGTCATGCAGAAATGCTGGACGTTATCGTGAAAAACGATATTAACCACTGGCAG
      GAGTGCTTCATTAGCGACCTAAAGCAGATAGTTCCGCGAAGCGCGGAAAGCCAGCAGCGCGATAAAGTTGCTAC
      CTTTCCAAAGCTTGCGTAG

Forward: GCTCTAGATGAGTCGTTTAGTCGTAG       52.4C(without) and 63.2(with)
Backward: GGACTAGTACTACGCAAGCTTTGGAAAG       54.5C(without) and 65.1(with)

Strain to use:BL21(DE3)
PAH Sequence:

      ATGTCCACTGCGGTCCTGGAAAACCCAGGCTTGGGCAGGAAACTCTCTGACTTTGGACAGGAAACAAGCTATAT
      TGAAGACAACTGCAATCAAAATGGTGCCATATCACTGATCTTCTCACTCAAAGAAGAAGTTGGTGCATTGGCCA
      AAGTATTGCGCTTATTTGAGGAGAATGATGTAAACCTGACCCACATTGAATCTAGACCTTCTCGTTTAAAGAAA
      GATGAGTATGAATTTTTCACCCATTTGGATAAACGTAGCCTGCCTGCTCTGACAAACATCATCAAGATCTTGAG
      GCATGACATTGGTGCCACTGTCCATGAGCTTTCACGAGATAAGAAGAAAGACACAGTGCCCTGGTTCCCAAGAA
      CCATTCAAGAGCTGGACAGATTTGCCAATCAGATTCTCAGCTATGGAGCGGAACTGGATGCTGACCACCCTGGT
      TTTAAAGATCCTGTGTACCGTGCAAGACGGAAGCAGTTTGCTGACATTGCCTACAACTACCGCCATGGGCAGCC
      CATCCCTCGAGTGGAATACATGGAGGAAGAAAAGAAAACATGGGGCACAGTGTTCAAGACTCTGAAGTCCTTGT
      ATAAAACCCATGCTTGCTATGAGTACAATCACATTTTTCCACTTCTTGAAAAGTACTGTGGCTTCCATGAAGAT
      AACATTCCCCAGCTGGAAGACGTTTCTCAATTCCTGCAGACTTGCACTGGTTTCCGCCTCCGACCTGTGGCTGG
      CCTGCTTTCCTCTCGGGATTTCTTGGGTGGCCTGGCCTTCCGAGTCTTCCACTGCACACAGTACATCAGACATG
      GATCCAAGCCCATGTATACCCCCGAACCTGACATCTGCCATGAGCTGTTGGGACATGTGCCCTTGTTTTCAGAT
      CGCAGCTTTGCCCAGTTTTCCCAGGAAATTGGCCTTGCCTCTCTGGGTGCACCTGATGAATACATTGAAAAGCT
      CGCCACAATTTACTGGTTTACTGTGGAGTTTGGGCTCTGCAAACAAGGAGACTCCATAAAGGCATATGGTGCTG
      GGCTCCTGTCATCCTTTGGTGAATTACAGTACTGCTTATCAGAGAAGCCAAAGCTTCTCCCCCTGGAGCTGGAG
      AAGACAGCCATCCAAAATTACACTGTCACGGAGTTCCAGCCCCTGTATTACGTGGCAGAGAGTTTTAATGATGC
      CAAGGAGAAAGTAAGGAACTTTGCTGCCACAATACCTCGGCCCTTCTCAGTTCGCTACGACCCATACACCCAAA
      GGATTGAGGTCTTGGACAATACCCAGCAGCTTAAGATTTTGGCTGATTCCATTAACAGTGAAATTGGAATCCTT
      TGCAGTGCCCTCCAGAAAATAAAGTAA

Forward: GCTCTAGATGTCCACTGCGGTCC       54.3C (without) & 66.0C (with)
Backward: GGACTAGTATTACTTTATTTTCTGGAGGGC       54.0C (without) & 64.0C (with)


Cellulase Sequence:

      ATGAAAAAAATTGTTTCTTTGGTTTGTGTGCTTGTGATGCTGGTAAGCATCTTAGGCTCGTTTTCAGTCGTA 
      GCGGCATCACCGGTAAAAGGCTTTCAGGTATCGGGAACAAAGCTTTTGGATGCAAGCGGAAACGAGCTTGTA 
      ATGAGGGGCATGCGTGATATTTCAGCAATAGATTTGGTTAAAGAAATAAAAATCGGATGGAATTTGGGAAAT 
      ACTTTGGATGCTCCTACAGAGACTGCCTGGGGAAATCCAAGGACAACCAAGGCAATGATAGAAAAGGTAAGG 
      GAAATGGGCTTTAATGCCGTCAGAGTGCCTGTTACCTGGGATACGCACATCGGACCTGCTCCGGACTATAAA 
      ATTGACGAAGCATGGCTGAACAGAGTTGAGGAAGTGGTAAACTATGTTCTTGACTGCGGTATGTACGCGATC 
      ATAAATGTTCACCATGACAATACATGGATTATACCTACATATGCCAATGAGCAAAGGAGTAAAGAAAAACTT 
      GTAAAAGTTTGGGAACAAATAGCAACCCGTTTTAAAGATTATGACGACCATTTGTTGTTTGAGACAATGAAC 
      GAACCGAGAGAAGTAGGTTCACCTATGGAATGGATGGGCGGAACGTATGAAAACCGAGATGTGATAAACAGA
      TTTAATTTGGCGGTTGTTAATACCATCAGAGCAAGCGGCGGAAATAACGATAAAAGATTCATACTGGTTCCG 
      ACCAATGCGGCAACCGGCCTGGATGTTGCATTAAACGACCTTGTCATTCCGAACAATGACAGCAGAGTCATA 
      GTATCCATACATGCTTATTCACCGTATTTCTTTGCTATGGATGTCAACGGAACTTCATATTGGGGAAGTGAC 
      TATGACAAGGCTTCTCTTACAAGTGAACTTGATGCTATTTACAACAGATTTGTGAAAAACGGAAGGGCTGTA 
      ATTATCGGAGAATTCGGAACCATTGACAAGAACAACCTGTCTTCAAGGGTGGCTCATGCCGAGCACTATGCA 
      AGAGAAGCAGTTTCAAGAGGAATTGCTGTTTTCTGGTGGGATAACGGCTATTACAATCCGGGTGATGCAGAG 
      ACTTATGCATTGCTGAACAGAAAAACTCTCTCATGGTATTATCCTGAAATTGTCCAGGCTCTTATGAGAGGT 
      GCCGGCGTTGAACCTTTAGTTTCACCGACTCCTACACCTACATTAATGCCGACCCCCTCGCCCACGGTGACA 
      GCAAATATTTTGTACGGTGACGTAAACGGGGACGGAAAAATAAATTCTACAGACTGTACAATGCTAAAGAGA 
      TATATTTTGCGTGGCATAGAAGAATTCCCAAGTCCTAGCGGAATTATAGCCGCTGACGTAAATGCGGATCTG 
      AAAATCAATTCCACCGACTTGGTATTGATGAAAAAATATCTACTGCGCTCAATAGACAAATTTCCTGCGGAG 
      GATTCTCAAACACCTGATGAAGACAATCCGGGCATTTTGTATAACGGAAGATTCGATTTTTCAGATCCGAAC 
      GGTCCGAAATGCGCCTGGTCCGGCAGCAATGTTGAGCTGAATTTTTACGGCACGGAAGCAAGTGTGACTATC
      AAATCCGGCGGTGAGAACTGGTTCCAGGCTATTGTAGACGGCAATCCTCTTCCTCCTTTTTCGGTTAACGCT
      ACTACCTCTACCGTAAAGCTTGTAAGCGGTCTTGCAGAAGGAGCTCATCATCTTGTATTGTGGAAGAGGACA
      GAGGCATCCTTGGGAGAAGTTCAGTTCCTTGGGTTTGATTTTGGTTCAGGAAAGCTTCTTGCCGCACCGAAG
      CCTTTGGAAAGAAAGATTGAGTTTATCGGAGACTCCATCACATGTGCATACGGAAATGAAGGAACAAGCAAG
      GAGCAGTCTTTTACACCGAAAAATGAAAACAGCTATATGTCTTATGCGGCAATTACAGCCCGTAATTTGAAT
      GCAAGTGCAAATATGATTGCGTGGTCCGGAATCGGACTTACCATGAACTACGGCGGAGCCCCCGGACCTCTT
      ATAATGGACCGTTATCCTTATACCCTTCCTTACAGCGGAGTCAGATGGGATTTTAGCAAATATGTGCCTCAG
      GTTGTTGTAATCAATCTTGGTACCAATGATTTTTCTACATCATTTGCAGATAAAACAAAGTTTGTAACGGCA
      TATAAAAACCTTATAAGTGAAGTTCGCAGGAACTATCCGGATGCCCATATATTCTGCTGTGTCGGTCCGATG
      CTTTGGGGAACGGGCCTGGATTTGTGCCGCAGTTATGTTACGGAAGTTGTAAATGATTGTAACAGAAGCGGG
      GATTTAAAGGTGTATTTTGTTGAGTTTCCGCAGCAGGACGGAAGCACCGGATACGGAGAAGACTGGCATCCA
      AGTATTGCCACCCACCAGCTGATGGCTGAGCGGCTTACTGCGGAAATAAAAAACAAGCTTGGATGGTAA


Forward: GCTCTAGATGAAAAAAATTGTTTCTTTGGTTTG       53.8C (without) & 62.0C (with)
Backward: GGACTAGTATTACCATCCAAGCTTGTTTTTTATTTC       57.4C (without) & 64.9C (with)

Dam Sequence:

      ATGAAGAAAAATCGCGCTTTTTTGAAGTGGGCAGGGGGCAAGTATCCCCTGCTTGATGATATTAAA
      CGGCATTTGCCCAAGGGCGAATGTCTGGTTGAGCCTTTTGTAGGTGCCGGGTCGGTGTTTCTCAAC
      ACCGACTTTTCTCGTTATATCCTTGCCGATATCAATAGCGACCTGATCAGTCTCTATAACATTGTG
      AAGATGCGTACTGATGAGTACGTACAGGCCGCACGCGAGCTGTTTGTTCCCGAAACAAATTGCGCC
      GAGGTTTACTATCAGTTCCGCGAAGAGTTCAACAAAAGCCAGGATCCGTTCCGTCGGGCGGTACTG
      TTTTTATATTTGAACCGCTACGGTTACAACGGCCTGTGTCGTTACAATCTGCGCGGTGAGTTTAAC
      GTGCCGTTCGGCCGCTACAAAAAACCCTATTTCCCGGAAGCAGAGTTGTATCACTTCGCTGAAAAA
      GCGCAGAATGCCTTTTTCTATTGTGAGTCTTACGCCGATAGCATGGCGCGCGCAGATGATGCATCC
      GTCGTCTATTGCGATCCGCCTTATGCACCGCTGTCTGCGACCGCCAACTTTACGGCGTATCACACA
      AACAGTTTTACGCTTGAACAACAAGCGCATCTGGCGGAGATCGCCGAAGGTCTGGTTGAGCGCCAT
      ATTCCAGTGCTGATCTCCAATCACGATACGATGTTAACGCGTGAGTGGTATCAGCGCGCAAAATTG
      CATGTCGTCAAAGTTCGACGCAGTATAAGCAGCAACGGCGGCACACGTAAAAAGGTGGACGAACTG
      CTGGCTTTGTACAAACCAGGAGTCGTTTCACCCGCGAAAAAATAA


Forward: GCTCTAGATGAAGAAAAATCGCGCTTTTTTG       55.9C (without) & 64.2C (with)
Backward: GGACTAGTATTATTTTTTCGCGGGTGAAAC       54.0C (without) & 64.0C (with)


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