BBa_J33205 1 BBa_J33205 hybrid lambda PR-lac promoter, repressed by lambda cI and lacI, plus lacZ 2006-10-25T11:00:00Z 2015-08-31T04:08:46Z The lambda-derived portion (bases 4 to 78) is derived from lambda DNA by PCR using primers based on published sequence (Genbank accession J02459, gi:215104). The lac portion (bases 84 to 353) is derived from E. coli BL21 genomic DNA using primers based on published sequence (Genbank accession J01636, gi:146575). Released HQ 2013 This part is a hybrid promoter designed to be repressed by either lambda cI or LacI. It consists of the the PRM-PR promoter region of bacteriophage lambda, including cI binding sites OR1, OR2 and OR3 as well as the -35 and -10 sites of both PRM and PR, fused via an introduced XhoI site to the LacI binding site of the lac promoter. (In this version, the lacZ' gene encoding the N-terminal 76 amino acid residues of LacZ has also been included to facilitate testing.) This approximately preserves the relationship between the -10 promoter region and and the LacI binding site. Our hope is that this promoter will be effectively repressed in the presence of either lambda cI or LacI. However, this has not been properly tested at the time of writing. We can say that good LacZ activity is observed in the presence of IPTG, indicating that the basic promoter function of lambda PR appears to be intact. Our sequencing also suggests two single nucleotide mutations within OR2 as compared to published sequence, which may be natural variants or PCR-induced mutations; in the latter case, they may or may not have some effect on cI binding. Note also that this construct includes the PRM promoter, which ought to be functional in the reverse direction. false false _63_ 0 837 63 In stock true This part includes the entire lambda PRM-PR region, with cI binding sites OR1, OR2 and OR3 and the -35 and -10 sequences of both promoters (PRM and PR). An XhoI site was introduced by PCR, and the DNA was joined to the region of the lac promoter beginning with the LacI binding site. The distance between the LacI binding site and the PR -10 site in the fusion is approximately the same as that between the LacI binding site and its native -10 site; thus we hope that LacI will be able to repress PR in our construct. However, this has yet to be properly tested. true Chris French annotation1905280 1 PR -10 range1905280 1 73 78 annotation1905267 1 cI binding site OR2 range1905267 1 36 52 annotation1905266 1 PRM -35 range1905266 1 32 37 annotation1905310 1 lacZ' range1905310 1 123 353 annotation1905309 1 rbs range1905309 1 112 115 annotation1905248 1 PRM -10 range1905248 1 9 14 annotation1905308 1 LacI binding site range1905308 1 85 105 annotation1905291 1 XhoI fusion site range1905291 1 79 84 annotation1905278 1 PR -35 range1905278 1 50 55 annotation1905279 1 cI binding site OR1 range1905279 1 60 76 annotation1905249 1 cI binding site OR3 range1905249 1 13 29 BBa_J33205_sequence 1 ctccgttaaatctatcaccgcaagggataaatatctaacaacgtgcgcgttgactattttacctctggcggtgataatctcgagaattgtgagcggataacaatttcacacaggaaacagctatgaccatgattacggattcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggcgctttgcctggtttccggcaccagaagcggtgccggaaagctggctggagtga igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 James Alastair McLaughlin Chris J. Myers 2017-03-06T15:00:00.000Z