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Wiki/Team:Warsaw/primers
From 2009.igem.org
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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 5' CTTGACCCGCAGTTGCAAAC 3' pZCseqR 5' CCCAACTGATCTTCAGCATC 3'
Sequencing of the T7 polymerase gene
T7-2seq 5' AGGTGCGCTGGCTGGCATC 3' T7-3seq 5' AAGGAAGGTTACTACTGGCT 3' T7-4seq 5' CAAAGAGTTCGGCTTCCGTC 3' T7Rseq 5' GCTTCAACCGCAGCTACCAC 3'
'Hunter and prey' strain preparation
Fusion of protein A and ompA and alpha or omega part of beta-lactamase
alphaL_link 5' CAGTGGTGGAAGCGGTGGCGGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGTCACCCAGAAACGCTGGTGAAAG 3' alphaP_XB 5' GG GGATCCGGTCTAGATTATTCGCCAGTTAATAGTTTGCG 3' omegaL_link 5' CAGTGGTGGAAGCGGTGGCGGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGTCTACTTACTCTAGCTTCCCGG 3' omegaP_EPB 5' AAGGATCCAACTGCAGAAGAATTCTTACCAATGCTTAATCAGTG 3' ompaL_N 5' CATATGAAAGCTACTAAACTGGTACTGGG 3' ompaP_link 5' GCCACCGCTTCCACCACTGCGGCCGCCAGAGCTCCCTCCTCCAGAACCTCCGCCGTTGTCCGGACGAGTGCCGATG 3'
Cloning of A and/or Z protein to expression vector (pET15b/pET24a)
AlphaP+link+Sac 5' GG GAGCTC CC ACCACCCCCACCCCCGCTACCTC 3' AlphaL+Nde 5' CGGG CATATG CACCCAGAAACGCTGGTGAAAG 3'
Mutagenesis of the A protein
ADelL+KpnI 5' AGGTACCCATGTAAGATCTCATAGAACGCG 3' AdelP 5' AACTTAAACGAAGAACAACGAAACGCC 3' AmutL+KpnI 5' TCGTTTAAGTTAGGTACCGGTGAGATCTCAGAGGACGCGTTTTGTTGTTCTTTGTTG 3' AmutP 5' AGAACAAGGAAACGCCTTCGCCCAAAGTTTAGCAGATGACCCAAGCCAAAGCGC 3'
Fusion of the A protein with the OmpaA protein and the alpha/omega part of beta-lactamase protein
AL+link10+homo2 5' TCTGGAGGTGGAGGTAGCGGGGGTGGGGGTGGTTCGGGTGGAGGTGGTAAAACCGCGGCTCTTGCGC 3' AlphaP+link10+homo2 5' CCGCTACCTCCACCTCCAGAACCACCTCCTCCGCTTCCTCCGCCACCTTCGCCAGTTAATAGTTTGCG 3' OmegaP+link10+homo2 5' CCGCTACCTCCACCTCCAGAACCACCTCCTCCGCTTCCTCCGCCACCCCAATGCTTAATCAGTGAGGC 3' AL+SacI 5' GG GAGCTCTAAAACCGCGGCTCTTGCGCAAC 3' AP+NotI 5' AA GCGGCCGCCGGTTGACTTCCCCGCGGAATTC 3' AlphaL+SacI 5' GG GAGCTCTCACCCAGAAACGCTGGTGAAAG 3' OmegaL+SacI 5' GGGAGCTCGCTACTTACTCTAGCTTCCCGG 3' AlphaPlinkSac 5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC 3'
Preparation of the BioBricks Standard Parts
AIDP_SpeNotPst 5' AAACTGCAGCGGCCGCTACTAGTATCAAAGTCCCAAAGTACGAAATG 3' AL_BNXNE 5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC 3' AlphaPSpe 5' GCACTAGTATTATTCGCCAGTTAATAGTTTGCG 3' APSacSpe 5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC 3' LinLSXNE 5' ATGAATTCGCGGCCGCTTCTAGAGGGAGCTCTGGTGGAAGCGGTGGCGGAGGATC 3' LinP_BS 5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC 3' LinP_BS2 5' ATACTAGTACCGGATCCAGAACCTCCTCCACCGCTACCTCCACCTCCAGAAC 3' LinPBSNP 5' AGCCTGCAGCGGCCGCTACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTACC 3' OmegPSpe 5' GCACTAGTATTACCAATGCTTAATCAGTGAG 3' OmegLNde 5' CAGCCATATGCTACTTACTCTAGCTTCCCGG 3' OmpLNXNE 5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC 3' OmLNXNEB 5' ATTAGATCTGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC 3' pETt7L_XNE 5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAATTAATACG 3' PlacL_XNE 5' TAGAATTCGCGGCCGCTTCTAGAGCTGGCACGACAGGTTTCCC 3' AP+0XhoI+SpeI 5' AAACTAGTCTCGAGGGTTGACTTCCCCGCGGAATTC 3' AIDl 5' TATCTAGAACCATGGACAGCCTCTTG 3' AraCl 5' GCTCTAGATTATGACAACTTGACGGC 3'
