Part:BBa_J33205

hybrid lambda PR-lac promoter, repressed by lambda cI and lacI, plus lacZ

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.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 79
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]