Wiki/Team:Warsaw/primers

Primers

'One-armed bandit' strain preparation

AID and T7 cloning T7lLinkB  5' CGGATCCGGCGGTGGCGGTAGTGGAGGAGGTGGGTCTGGAGGCGGAGGGAACACGATTAACATCGCTAAGAAC  3' T7lRBSHi  5' CACAAGCTTAGGAGGTGTACTAGATGAACACGATTAACATCGCTAAG  3' T7pXbSal  5' TCTGTCGACTCTAGATTACGCGAACGCGAAGTCC  3' AIDlNrH   5' CGTAAGCTTAGGAGGTGTAGACCATGGACAGCCTCTTGATG  3' AIDpLinB  5' CCACTACCGCCACCGCCGGATCCGCCACCTCCGCTGCCTCCACCACCAAGTCCCAAAGTACGAAATGC  3'

Sequencing of pMPM-T5 pMPMT5seqF 5' CTACTGTTTCTCCATACCCG  3' pMPMT5seqR 5' GAAAATCTTCTCTCATCCGC 3' AIDseqP    5' GCTTCGGCGCATCCTTTTG 3'

Sequencing of β-galactosidase from pZC320 pZCseqL</a>    5' CTTGACCCGCAGTTGCAAAC 3' pZCseqR</a>    5' CCCAACTGATCTTCAGCATC 3'

Sequencing of the T7 polymerase gene T7-2seq</a>  5' AGGTGCGCTGGCTGGCATC 3' T7-3seq</a>  5' AAGGAAGGTTACTACTGGCT 3' T7-4seq</a>  5' CAAAGAGTTCGGCTTCCGTC 3' T7Rseq</a>   5' GCTTCAACCGCAGCTACCAC 3'

'Hunter and prey' strain preparation

Fusion of protein A and ompA and alpha or omega part of beta-lactamase alphaL_link</a> 5' CAGTGGTGGAAGCGGTGGCGGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGTCACCCAGAAACGCTGGTGAAAG 3' alphaP_XB</a>   5' GG GGATCCGGTCTAGATTATTCGCCAGTTAATAGTTTGCG 3' omegaL_link</a> 5' CAGTGGTGGAAGCGGTGGCGGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGTCTACTTACTCTAGCTTCCCGG 3' omegaP_EPB</a>  5' AAGGATCCAACTGCAGAAGAATTCTTACCAATGCTTAATCAGTG 3' ompaL_N</a>     5' CATATGAAAGCTACTAAACTGGTACTGGG 3' ompaP_link</a>  5' GCCACCGCTTCCACCACTGCGGCCGCCAGAGCTCCCTCCTCCAGAACCTCCGCCGTTGTCCGGACGAGTGCCGATG 3'

Cloning of A and/or Z protein to expression vector (pET15b/pET24a) AlphaP+link+Sac</a>  5' GG GAGCTC CC ACCACCCCCACCCCCGCTACCTC  3' AlphaL+Nde</a>       5' CGGG CATATG CACCCAGAAACGCTGGTGAAAG  3'

Mutagenesis of the A protein ADelL+KpnI</a> 5' AGGTACCCATGTAAGATCTCATAGAACGCG  3' AdelP</a>      5' AACTTAAACGAAGAACAACGAAACGCC  3' <a name="AmutL+KpnI">AmutL+KpnI</a> 5' TCGTTTAAGTTAGGTACCGGTGAGATCTCAGAGGACGCGTTTTGTTGTTCTTTGTTG  3' <a name="AmutP">AmutP</a>      5' AGAACAAGGAAACGCCTTCGCCCAAAGTTTAGCAGATGACCCAAGCCAAAGCGC  3'

Fusion of the A protein with the OmpaA protein and the alpha/omega part of beta-lactamase protein <a name="AL+link10+homo2">AL+link10+homo2</a>    5' TCTGGAGGTGGAGGTAGCGGGGGTGGGGGTGGTTCGGGTGGAGGTGGTAAAACCGCGGCTCTTGCGC  3' <a name="AlphaP+link10+homo2">AlphaP+link10+homo2</a> 5' CCGCTACCTCCACCTCCAGAACCACCTCCTCCGCTTCCTCCGCCACCTTCGCCAGTTAATAGTTTGCG 3' <a name="OmegaP+link10+homo2">OmegaP+link10+homo2</a> 5' CCGCTACCTCCACCTCCAGAACCACCTCCTCCGCTTCCTCCGCCACCCCAATGCTTAATCAGTGAGGC 3' <a name="AL+SacI">AL+SacI</a>            5' GG GAGCTCTAAAACCGCGGCTCTTGCGCAAC  3' <a name="AP+NotI">AP+NotI</a>            5' AA GCGGCCGCCGGTTGACTTCCCCGCGGAATTC  3' <a name="AlphaL+SacI">AlphaL+SacI</a>        5' GG GAGCTCTCACCCAGAAACGCTGGTGAAAG  3' <a name="OmegaL+SacI">OmegaL+SacI</a>        5' GGGAGCTCGCTACTTACTCTAGCTTCCCGG  3' <a name="AlphaPlinkSac">AlphaPlinkSac</a>      5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC  3'

Preparation of the BioBricks Standard Parts <a name="AIDpSNP">AIDP_SpeNotPst</a>      5' AAACTGCAGCGGCCGCTACTAGTATCAAAGTCCCAAAGTACGAAATG  3' <a name="AL_BNXNE">AL_BNXNE</a>            5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC  3' <a name="AlphaPSpe">AlphaPSpe</a>           5' GCACTAGTATTATTCGCCAGTTAATAGTTTGCG  3' <a name="APSacSpe">APSacSpe</a>            5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC  3' <a name="LinLSXNE">LinLSXNE</a>            5' ATGAATTCGCGGCCGCTTCTAGAGGGAGCTCTGGTGGAAGCGGTGGCGGAGGATC  3' <a name="LinP_BS">LinP_BS</a>             5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC  3' <a name="LinP_BS2">LinP_BS2</a>            5' ATACTAGTACCGGATCCAGAACCTCCTCCACCGCTACCTCCACCTCCAGAAC  3' <a name="LinPBSNP">LinPBSNP</a>            5' AGCCTGCAGCGGCCGCTACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTACC  3' <a name="OmegPSpe">OmegPSpe</a>            5' GCACTAGTATTACCAATGCTTAATCAGTGAG  3' <a name="OmegLNde">OmegLNde</a>            5' CAGCCATATGCTACTTACTCTAGCTTCCCGG  3' <a name="OmpLNXNE">OmpLNXNE</a>            5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC  3' <a name="OmLNXNEB">OmLNXNEB</a>            5' ATTAGATCTGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC  3' <a name="PETt7L_XNE">pETt7L_XNE</a>          5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAATTAATACG  3' <a name="PlacL_XNE">PlacL_XNE</a>           5' TAGAATTCGCGGCCGCTTCTAGAGCTGGCACGACAGGTTTCCC  3' <a name="AP+XhoI+SpeI">AP+0XhoI+SpeI</a>       5' AAACTAGTCTCGAGGGTTGACTTCCCCGCGGAATTC  3' <a name=AIDl>AIDl</a>                5' TATCTAGAACCATGGACAGCCTCTTG 3' <a name=AraCl>AraCl</a>               5' GCTCTAGATTATGACAACTTGACGGC 3'