IGEM:IMPERIAL/2009/Primers: Difference between revisions
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http://www.premierbiosoft.com/tech_notes/PCR_Primer_Design.html | 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.</b> | <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> | ||
| Line 21: | Line 21: | ||
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> | ||
<br> | <br> | ||
<b>OtsB Sequence:</b> <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> | ||
<br> | <br> | ||
<b>B3023 Sequence:</b><br> | <b>B3023 Sequence:</b><br> | ||
| Line 57: | Line 63: | ||
<br> | <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> | <br> | ||
<b>WaaL Sequence:</b><br> | <b>WaaL Sequence:</b><br> | ||
| Line 88: | Line 95: | ||
<br> | <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> | ||
<br> | <br> | ||
| Line 119: | Line 126: | ||
CTTTCCAAAGCTTGCGTAG | 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> | ||
<br> | <br> | ||
| Line 151: | Line 158: | ||
Forward: <b>GCTCTAG</b>ATGTCCACTGCGGTCC 54.3C (without) & 66.0C (with)<br> | Forward: <b>GCTCTAG</b>ATGTCCACTGCGGTCC 54.3C (without) & 66.0C (with)<br> | ||
Backward: <b>GGACTAGTA</b>TTACTTTATTTTCTGGAGGGC 54.0C (without) & 64.0C (with)<br> | Backward: <b>GGACTAGTA</b>TTACTTTATTTTCTGGAGGGC 54.0C (without) & 64.0C (with)<br> | ||
<br> | |||
<b>Sequence Length:</b> 1359bp | |||
<br> | <br> | ||
<br> | <br> | ||
| Line 193: | Line 202: | ||
Forward: <b>GCTCTAG</b>ATGAAAAAAATTGTTTCTTTGGTTTG 53.8C (without) & 62.0C (with)<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> | 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> | <br> | ||
Latest revision as of 02:24, 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)
