Part:BBa_K1132016

Promoter (OmpR, positive) followed by RFP protein generator and Cph8 (Cph1/EnvZ fusion) under TetR r

Chimeric Cph1 light receptor/EnvZ protein requires phycocyanobilin (PCB) biosynthetic genes for PCB formation (ho1 and PcyA). The exposure to red light (660nm) inhibits the activity of the EnvZ histidine kinase domain. In the dark, the EnvZ histidine kinase phosphorylates endogenous OmpR, a transcription factor which activates transcription from the OmpC promoter. This part must be used in E.coli deficient in wild-type EnvZ.

The promoter p(TetR) is constitutively on and repressed by TetR (Part:BBa_P0440). TetR binds to the p(TetR) and inhibits its operation. It regulates Cph8 (Part:BBa_K1132015) production. Tetracycline or aTc (anhydrotetracycline) binds to TetR and allows Cph8 production.

This part is an intermediate Biobrick for the characterisation of the red light responsive system (Part:BBa_K1132017) with the RFP protein as output. The monomeric RFP is a Red Fluorescent Protein with an excitation peak at 584 nm and an emission peak at 607 nm. The RFP is positively regulated by OmpR-controlled promoter. Phosphorylated OmpR binds to the operator sites and activates transcription of RFP.

This must be used with Part:BBa_K1132013 composed of the heme oxygenase (ho1), ferredoxin oxidoreductase (PcyA) and tetracycline repressor (TetR) under constitutive promoter.

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 1342
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 692
  Illegal AgeI site found at 804
  
• 1000
  COMPATIBLE WITH RFC[1000]