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+link14+homo (alphaL_link) 5' CAGTGGTGGAAGCGGTGGC GGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGT CACCCAGAAACGCTGGTGAAAG 3'
alphaP+STOP+XbaI+BamHI (alphaP_XB) 5' GG GGATCC GG TCTAGA TTA TTCGCCAGTTAATAGTTTGCG 3'
omegaL+link14+homo (omegaL_link) 5' CAGTGGTGGAAGCGGTGGC GGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGT CTACTTACTCTAGCTTCCCGG 3'
omegaP+STOP+EcoRI+PstI+BamHI (omegaP_EPB) 5' AAGGATCCAACTGCAGAAGAATTC TTACCAATGCTTAATCAGTG 3'
OmpAL+NdeI (ompaL_N) 5' CATATGAAAGCTACTAAACTGGTACTGGG 3'
OmpAP+link8+SacI+NotI+homo (ompaP_link) 5' GCCACCGCTTCCACCACT GCGGCCGC CA GAGCTC CCTCCTCCAGAACCTCC GCCGTTGTCCGGACGAGTGCCGATG 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' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC 3'
AlphaP+link10+homo2 5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC TTCGCCAGTTAATAGTTTGCG 3'
OmegaP+link10+homo2 5' CCGCTACCTCCACCTCCAGA ACCACCTCCTCCGCTTCCTCCGCCACC CCAATGCTTAATCAGTGAGGC 3'
AL+SacI 5' GG GAGCTC T AAAACCGCGGCTCTTGCGCAAC 3'
AP+NotI 5' AA GCGGCCGC C GGTTGACTTCCCCGCGGAATTC 3'
AlphaL+SacI 5' GG GAGCTC T CACCCAGAAACGCTGGTGAAAG 3'
OmegaL+SacI 5' GG GAGCTC G CTACTTACTCTAGCTTCCCGG 3'
AlphaPlinkSac 5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC 3'
Preparation of the BioBricks Standard Parts
AIDP_HindSpeNotPst 5' AAACTGCAGCGGCCGCTACTAGTAAAGCTTTCAAAGTCCCAAAGTACGAAATG 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' AA ACTAGT CTCGAG GGTTGACTTCCCCGCGGAATTC 3'
AIDl 5' TATCTAGAACCATGGACAGCCTCTTG 3'
AraCl 5' GCTCTAGATTATGACAACTTGACGGC 3'
|