IGEM:IMPERIAL/2009/Primers: Difference between revisions
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=Primers to be used for PCR= | =Primers to be used for PCR= | ||
Considerations for primer design | |||
http://www.premierbiosoft.com/tech_notes/PCR_Primer_Design.html | |||
<b>Use [http://www.eurofinsdna.com/de/home.html?ad=Brand&gclid=CIPQ3f3Jh5wCFZ0U4wodG1b7_w this] website to calculate temperatures and modify primers and [http://www.bioinformatics.org/sms/rev_comp.html this] one for reverse complement.</b> | |||
<b>RcsB Sequence:</b> | <b>RcsB Sequence:</b> | ||
ATGAACAATATGAACGTAATTATTGCCGATGACCATCCGATAGTCTTGTTCGGTATTCGCAAATCACTTG | ATGAACAATATGAACGTAATTATTGCCGATGACCATCCGATAGTCTTGTTCGGTATTCGCAAATCACTTG | ||
AGCAAATTGAGTGGGTGAATGTTGTCGGCGAATTTGAAGACTCTACAGCACTGATCAACAACCTGCCGAA | AGCAAATTGAGTGGGTGAATGTTGTCGGCGAATTTGAAGACTCTACAGCACTGATCAACAACCTGCCGAA | ||
ACTGGATGCGCATGTGTTGATTACCGATCTCTCCATGCCTGGCGATAAGTACGGCGATGGCATTACCTTA | ACTGGATGCGCATGTGTTGATTACCGATCTCTCCATGCCTGGCGATAAGTACGGCGATGGCATTACCTTA | ||
ATCAAGTACATCAAGCGCCATTTCCCAAGCCTGTCGATCATTGTTCTGACTATGAACAACAACCCGGCGA | ATCAAGTACATCAAGCGCCATTTCCCAAGCCTGTCGATCATTGTTCTGACTATGAACAACAACCCGGCGA | ||
TTCTTAGTGCGGTATTGGATCTGGATATCGAAGGGATCGTGCTGAAACAAGGTGCACCGACCGATCTGCC | TTCTTAGTGCGGTATTGGATCTGGATATCGAAGGGATCGTGCTGAAACAAGGTGCACCGACCGATCTGCC | ||
GAAAGCTCTCGCCGCGCTGCAGAAAGGGAAGAAATTTACCCCGGAAAGCGTTTCTCGCCTGTTGGAAAAA | GAAAGCTCTCGCCGCGCTGCAGAAAGGGAAGAAATTTACCCCGGAAAGCGTTTCTCGCCTGTTGGAAAAA | ||
ATCAGTGCTGGTGGTTACGGTGACAAGCGTCTCTCGCCAAAAGAGAGTGAAGTTCTGCGCCTGTTTGCGG | ATCAGTGCTGGTGGTTACGGTGACAAGCGTCTCTCGCCAAAAGAGAGTGAAGTTCTGCGCCTGTTTGCGG | ||
AAGGCTTCCTGGTGACCGAGATCGCTAAAAAGCTGAACCGCAGTATTAAAACCATCAGTAGCCAGAAGAA | AAGGCTTCCTGGTGACCGAGATCGCTAAAAAGCTGAACCGCAGTATTAAAACCATCAGTAGCCAGAAGAA | ||
ATCTGCGATGATGAAGCTGGGTGTCGAGAACGATATCGCCCTGCTGAATTATCTCTCTTCAGTGACCTTA | ATCTGCGATGATGAAGCTGGGTGTCGAGAACGATATCGCCCTGCTGAATTATCTCTCTTCAGTGACCTTA | ||
AGTCCGGCAGATAAAGACTAA | AGTCCGGCAGATAAAGACTAA | ||
Forward: <b>GCTCTAG</b>ATGAACAATATGAACGTAATTATTG 53.1C(without) and 61.8(with)<br> | |||
Backward: <b>GGACTAGTA</b>TTAGTCTTTATCTGCCGG 51.4C(without) and 63.4(with)<br> | |||
<br> | |||
<b>Sequence Length:</b> 651bp | |||
<br> | |||
Strains to use:DH10B/TOP10 | |||
<br> | |||
<br> | |||
<b>OtsB Sequence:</b> <br> | |||
ATGACAGAACCGTTAACCGAAACCCCTGAACTATCCGCGAAATATGCCTGGTTTTTTGATCTTGATG | |||
GAACGCTGGCGGAAATCAAACCGCATCCCGATCAGGTCGTCGTGCCTGACAATATTCTGCAAGGACT | |||
ACAGCTACTGGCAACCGCAAGTGATGGTGCATTGGCATTGATATCAGGGCGCTCAATGGTGGAGCTT | |||
GACGCACTGGCAAAACCTTATCGCTTCCCGTTAGCGGGCGTGCATGGGGCGGAGCGCCGTGACATCA | |||
ATGGTAAAACACATATCGTTCATCTGCCGGATGCGATTGCGCGTGATATTAGCGTGCAACTGCATAC | |||
AGTCATCGCTCAGTATCCCGGCGCGGAGCTGGAGGCGAAAGGGATGGCTTTTGCGCTGCATTATCGT | |||
CAGGCTCCGCAGCATGAAGACGCATTAATGACATTAGCGCAACGTATTACTCAGATCTGGCCACAAA | |||
TGGCGTTACAGCAGGGAAAGTGTGTTGTCGAGATCAAACCGAGAGGTACCAGTAAAGGTGAGGCAAT | |||
TGCAGCTTTTATGCAGGAAGCTCCCTTTATCGGGCGAACGCCCGTATTTCTGGGCGATGATTTAACC | |||
GATGAATCTGGCTTCGCAGTCGTTAACCGACTGGGCGGAATGTCAGTAAAAATTGGCACAGGTGCAA | |||
CTCAGGCATCATGGCGACTGGCGGGTGTGCCGGATGTCTGGAGCTGGCTTGAAATGATAACCACCGC | |||
ATTACAACAAAAAAGAGAAAATAACAGGAGTGATGACTATGAGTCGTTTAGTCGTAGTATCTAA | |||
<br> | |||
Forward: <b>GCTCTAG</b>ATGACAGAACCGTTAACCG 54.5C (without) & 64.8C (with)<br> | |||
Backward: <b>GGACTAGTA</b>TTAGATACTACGACTAAACGAC 54.7C (without) & 64.2C (with)<br> | |||
<br> | |||
<b>Sequence Length:</b> 805bp | |||
<br> | |||
Strains to use:BL21(DE3) | |||
<br> | |||
<br> | |||
<b>B3023 Sequence:</b><br> | |||
ATGACAAACCTGACACTGGATGTAAACATTATCGATTTCCCATCAATACCTGTGGCGATGTTGCCG | |||
CACCGCTGTAGCCCTGAATTGCTCAACTACAGCGTGGCGAAATTTATCATGTGGCGTAAAGAGACG | |||
GGGCTTTCTCCTGTTAACCAAAGCCAGACTTTTGGCGTCGCCTGGGACGACCCTGCCACCACCGCA | |||
CCGGAAGCGTTTCGCTTTGATATCTGCGGCAGCGTTAGCGAACCGATTCCCGATAATCGTTATGGT | |||
GTGAGCAATGGTGAACTTACCGGTGGACGTTATGCCGTGGCCCGCCACGTTGGCGAGCTGGACGAT | |||
ATTTCACACACGGTATGGGGCATCATTCGCCACTGGCTGCCTGCAAGCGGCGAGAAAATGCGTAAA | |||
GCACCGATTCTGTTTCACTACACCAATCTTGCCGAAGGGGTGACAGAGCAGCGACTGGAAACGGAT | |||
GTTTATGTGCCGTTGGCGTGA | |||
<br> | |||
Forward: <b>GCTCTAG</b>ATGACAAACCTGACACTG 51.4C(without) and 63.0(with)<br> | |||
Backward: <b>GGACTAGTA</b>TCACGCCAACGGCAC 53.3C(without) and 66.1C(with)<br> | |||
<br> | |||
<b>Sequence Length:</b> 483bp | |||
<br> | |||
Strains to use:BL21(DE3) | |||
<br> | |||
<br> | |||
<b>WaaL Sequence:</b><br> | |||
ATGCTAACATCCTTTAAACTTCATTCATTGAAACCTTACACTCTGAAATCATCAATGATTTTAGAG | |||
ATAATAACTTATATATTATGTTTTTTTTCAATGATAATTGCATTCGTCGATAATACTTTCAGCATA | |||
AAAATATATAATATCACTGCTATAGTTTGCTTATTGTCACTAATTTTACGTGGCAGACAAGAAAATT | |||
ATAATATAAAAAACCTTATTCTTCCCCTTTCTATATTTTTAATAGGCTTGCTTGATTTAATTTGGTA | |||
TTCTGCGTTTAAAGTAGATAATTCGCCATTTCGTGCTACTTACCATAGTTATTTAAATACTGCCAAA | |||
ATATTTATATTTGGTTCTTTTATTGTTTTCTTGACACTAACTAGCCAGCTAAAATCAAAAAAAGAGA | |||
GTGTATTATACACTTTGTATTCTCTGTCATTTCTAATTGCTGGATATGCAATGTATATTAATAGCAT | |||
TCATGAAAATGACCGCATTTCTTTTGGTGTAGGAACGGCAACAGGAGCAGCATATTCAACAATGCTA | |||
ATAGGGATAGTTAGTGGCGTTGCGATTCTTTATACTAAGAAAAATCATCCTTTTTTATTTTTATTAA | |||
ATAGTTGCGCGGTACTTTATGTTCTGGCGCTAACACAAACCAGAGCAACCCTACTCCTGTTCCCTAT | |||
AATTTGTGTTGCTGCATTAATAGCTTATTATAATAAATCACCCAAGAAATTCACTTCCTCTATTGTT | |||
CTACTAATTGCTATATTAGCTAGCATTGTTATTATATTTAATAAACCAATACAGAATCGCTATAATG | |||
AAGCATTAAATGACTTAAACAGTTATACCAATGCTAATAGTGTTACTTCCCTAGGTGCAAGACTGGC | |||
AATGTACGAAATTGGTTTAAATATATTCATAAAGTCACCTTTTTCATTTAGATCAGCAGAGTCACGC | |||
GCTGAAAGTATGAATTTGTTAGTTGCAGAACACAATAGGCTAAGAGGGGCATTGGAGTTTTCTAACG | |||
TACATCTACATAATGAGATAATTGAAGCAGGGTCACTGAAAGGTCTGATGGGAATTTTTTCCACACT | |||
TTTCCTCTATTTTTCACTATTTTATATAGCATATAAAAAACGAGCTTTGGGTTTGTTGATATTAACG | |||
CTTGGCATTGTGGGGATTGGACTCAGTGATGTGATCATATGGGCACGCAGCATTCCAATTATCATTA | |||
TATCCGCTATAGTCCTCTTACTCGTCATTAATAATCGTAACAATACAATTAATTAA | |||
<br> | |||
Forward: <b>GCTCTAG</b>ATGCTAACATCCTTTAAACTTC 52.8C(without) and 62.4(with)<br> | |||
Backward: <b>GGACTAGTA</b>TTAATTAATTGTATTGTTACGATTATTAATG 54.9C(without) and 62.3(with)<br> | |||
<br> | |||
<b>Sequence Length:</b> 1260bp | |||
<br> | |||
Strains to use:DH10B/TOP10 | |||
<br> | |||
<br> | |||
<b>OtsA Sequence:</b> | |||
ATGAGTCGTTTAGTCGTAGTATCTAACCGGATTGCACCACCAGACGAGCACGCCGCCAGTGCCGGTGGCCTTGC | |||
CGTTGGCATACTGGGGGCACTGAAAGCCGCAGGCGGACTGTGGTTTGGCTGGAGTGGTGAAACAGGGAATGAGG | |||
ATCAGCCGCTAAAAAAGGTGAAAAAAGGTAACATTACGTGGGCCTCTTTTAACCTCAGCGAACAGGACCTTGAC | |||
GAATACTACAACCAATTCTCCAATGCCGTTCTCTGGCCCGCTTTTCATTATCGGCTCGATCTGGTGCAATTTCA | |||
GCGTCCTGCCTGGGACGGCTATCTACGCGTAAATGCGTTGCTGGCAGATAAATTACTGCCGCTGTTGCAAGACG | |||
ATGACATTATCTGGATCCACGATTATCACCTGTTGCCATTTGCGCATGAATTACGCAAACGGGGAGTGAATAAT | |||
CGCATTGGTTTCTTTCTGCATATTCCTTTCCCGACACCGGAAATCTTCAACGCGCTGCCGACATATGACACCTT | |||
GCTTGAACAGCTTTGTGATTATGATTTGCTGGGTTTCCAGACAGAAAACGATCGTCTGGCGTTCCTGGATTGTC | |||
TTTCTAACCTGACCCGCGTCACGACACGTAGCGCAAAAAGCCATACAGCCTGGGGCAAAGCATTTCGAACAGAA | |||
GTCTACCCGATCGGCATTGAACCGAAAGAAATAGCCAAACAGGCTGCCGGGCCACTGCCGCCAAAACTGGCGCA | |||
ACTTAAAGCGGAACTGAAAAACGTACAAAATATCTTTTCTGTCGAACGGCTGGATTATTCCAAAGGTTTGCCAG | |||
AGCGTTTTCTCGCCTATGAAGCGTTGCTGGAAAAATATCCGCAGCATCATGGTAAAATTCGTTATACCCAGATT | |||
GCACCAACGTCGCGTGGTGATGTGCAAGCCTATCAGGATATTCGTCATCAGCTCGAAAATGAAGCTGGACGAAT | |||
TAATGGTAAATACGGGCAATTAGGCTGGACGCCGCTTTATTATTTGAATCAGCATTTTGACCGTAAATTACTGA | |||
TGAAAATATTCCGCTACTCTGACGTGGGCTTAGTGACGCCACTGCGTGACGGGATGAACCTGGTAGCAAAAGAG | |||
TATGTTGCTGCTCAGGACCCAGCCAATCCGGGCGTTCTTGTTCTTTCGCAATTTGCGGGAGCGGCAAACGAGTT | |||
AACGTCGGCGTTAATTGTTAACCCCTACGATCGTGACGAAGTTGCAGCTGCGCTGGATCGTGCATTGACTATGT | |||
CGCTGGCGGAACGTATTTCCCGTCATGCAGAAATGCTGGACGTTATCGTGAAAAACGATATTAACCACTGGCAG | |||
GAGTGCTTCATTAGCGACCTAAAGCAGATAGTTCCGCGAAGCGCGGAAAGCCAGCAGCGCGATAAAGTTGCTAC | |||
CTTTCCAAAGCTTGCGTAG | |||
Forward: <b>GCTCTAG</b>ATGAGTCGTTTAGTCGTAG 52.4C(without) and 63.2(with)<br> | |||
Backward: <b>GGACTAGTA</b>CTACGCAAGCTTTGGAAAG 54.5C(without) and 65.1(with)<br> | |||
<br> | |||
<b>Sequence Length:</b> 1425bp | |||
<br> | |||
Strain to use:BL21(DE3) | |||
<br> | |||
<br> | |||
<b>PAH Sequence:</b> | |||
ATGTCCACTGCGGTCCTGGAAAACCCAGGCTTGGGCAGGAAACTCTCTGACTTTGGACAGGAAACAAGCTATAT | |||
TGAAGACAACTGCAATCAAAATGGTGCCATATCACTGATCTTCTCACTCAAAGAAGAAGTTGGTGCATTGGCCA | |||
AAGTATTGCGCTTATTTGAGGAGAATGATGTAAACCTGACCCACATTGAATCTAGACCTTCTCGTTTAAAGAAA | |||
GATGAGTATGAATTTTTCACCCATTTGGATAAACGTAGCCTGCCTGCTCTGACAAACATCATCAAGATCTTGAG | |||
GCATGACATTGGTGCCACTGTCCATGAGCTTTCACGAGATAAGAAGAAAGACACAGTGCCCTGGTTCCCAAGAA | |||
CCATTCAAGAGCTGGACAGATTTGCCAATCAGATTCTCAGCTATGGAGCGGAACTGGATGCTGACCACCCTGGT | |||
TTTAAAGATCCTGTGTACCGTGCAAGACGGAAGCAGTTTGCTGACATTGCCTACAACTACCGCCATGGGCAGCC | |||
CATCCCTCGAGTGGAATACATGGAGGAAGAAAAGAAAACATGGGGCACAGTGTTCAAGACTCTGAAGTCCTTGT | |||
ATAAAACCCATGCTTGCTATGAGTACAATCACATTTTTCCACTTCTTGAAAAGTACTGTGGCTTCCATGAAGAT | |||
AACATTCCCCAGCTGGAAGACGTTTCTCAATTCCTGCAGACTTGCACTGGTTTCCGCCTCCGACCTGTGGCTGG | |||
CCTGCTTTCCTCTCGGGATTTCTTGGGTGGCCTGGCCTTCCGAGTCTTCCACTGCACACAGTACATCAGACATG | |||
GATCCAAGCCCATGTATACCCCCGAACCTGACATCTGCCATGAGCTGTTGGGACATGTGCCCTTGTTTTCAGAT | |||
CGCAGCTTTGCCCAGTTTTCCCAGGAAATTGGCCTTGCCTCTCTGGGTGCACCTGATGAATACATTGAAAAGCT | |||
CGCCACAATTTACTGGTTTACTGTGGAGTTTGGGCTCTGCAAACAAGGAGACTCCATAAAGGCATATGGTGCTG | |||
GGCTCCTGTCATCCTTTGGTGAATTACAGTACTGCTTATCAGAGAAGCCAAAGCTTCTCCCCCTGGAGCTGGAG | |||
AAGACAGCCATCCAAAATTACACTGTCACGGAGTTCCAGCCCCTGTATTACGTGGCAGAGAGTTTTAATGATGC | |||
CAAGGAGAAAGTAAGGAACTTTGCTGCCACAATACCTCGGCCCTTCTCAGTTCGCTACGACCCATACACCCAAA | |||
GGATTGAGGTCTTGGACAATACCCAGCAGCTTAAGATTTTGGCTGATTCCATTAACAGTGAAATTGGAATCCTT | |||
TGCAGTGCCCTCCAGAAAATAAAGTAA | |||
Forward: <b>GCTCTAG</b>ATGTCCACTGCGGTCC 54.3C (without) & 66.0C (with)<br> | |||
Backward: <b>GGACTAGTA</b>TTACTTTATTTTCTGGAGGGC 54.0C (without) & 64.0C (with)<br> | |||
<br> | |||
<b>Sequence Length:</b> 1359bp | |||
<br> | |||
<br> | |||
<b>Cellulase Sequence:</b> | |||
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 | |||
<br> | |||
Forward: <b>GCTCTAG</b>ATGAAAAAAATTGTTTCTTTGGTTTG 53.8C (without) & 62.0C (with)<br> | |||
Backward: <b>GGACTAGTA</b>TTACCATCCAAGCTTGTTTTTTATTTC 57.4C (without) & 64.9C (with)<br> | |||
<br> | |||
<b>Sequence Length:</b> 2445bp | |||
<br> | |||
<br> | |||
<b>Dam Sequence:</b> | |||
ATGAAGAAAAATCGCGCTTTTTTGAAGTGGGCAGGGGGCAAGTATCCCCTGCTTGATGATATTAAA | |||
CGGCATTTGCCCAAGGGCGAATGTCTGGTTGAGCCTTTTGTAGGTGCCGGGTCGGTGTTTCTCAAC | |||
ACCGACTTTTCTCGTTATATCCTTGCCGATATCAATAGCGACCTGATCAGTCTCTATAACATTGTG | |||
AAGATGCGTACTGATGAGTACGTACAGGCCGCACGCGAGCTGTTTGTTCCCGAAACAAATTGCGCC | |||
GAGGTTTACTATCAGTTCCGCGAAGAGTTCAACAAAAGCCAGGATCCGTTCCGTCGGGCGGTACTG | |||
TTTTTATATTTGAACCGCTACGGTTACAACGGCCTGTGTCGTTACAATCTGCGCGGTGAGTTTAAC | |||
GTGCCGTTCGGCCGCTACAAAAAACCCTATTTCCCGGAAGCAGAGTTGTATCACTTCGCTGAAAAA | |||
GCGCAGAATGCCTTTTTCTATTGTGAGTCTTACGCCGATAGCATGGCGCGCGCAGATGATGCATCC | |||
GTCGTCTATTGCGATCCGCCTTATGCACCGCTGTCTGCGACCGCCAACTTTACGGCGTATCACACA | |||
AACAGTTTTACGCTTGAACAACAAGCGCATCTGGCGGAGATCGCCGAAGGTCTGGTTGAGCGCCAT | |||
ATTCCAGTGCTGATCTCCAATCACGATACGATGTTAACGCGTGAGTGGTATCAGCGCGCAAAATTG | |||
CATGTCGTCAAAGTTCGACGCAGTATAAGCAGCAACGGCGGCACACGTAAAAAGGTGGACGAACTG | |||
CTGGCTTTGTACAAACCAGGAGTCGTTTCACCCGCGAAAAAATAA | |||
<br> | |||
Forward: <b>GCTCTAG</b>ATGAAGAAAAATCGCGCTTTTTTG 55.9C (without) & 64.2C (with)<br> | |||
Backward: <b>GGACTAGTA</b>TTATTTTTTCGCGGGTGAAAC 54.0C (without) & 64.0C (with)<br> | |||
<br> | |||
<b>Sequence Length:</b> 837bp | |||
<br> | |||
Strain to use:BL21(DE3) | |||
<br> | |||
Latest revision as of 09:24, 12 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)
Sequence Length: 651bp
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)
Sequence Length: 805bp
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)
Sequence Length: 483bp
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)
Sequence Length: 1260bp
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)
Sequence Length: 1425bp
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)
Sequence Length: 1359bp
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)
Sequence Length: 2445bp
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)
Sequence Length: 837bp
Strain to use:BL21(DE3)
