Part:BBa_K4194017
pRM'-RBS-LacI-LVA-RBS-mRFP1-LVA-terminator
The expression of mRFP1 and LacI is regulated by promoter pRM', which can be specifically activated by protein CI. The mRFP1 was used as a reporter gene.
Promoter pRM' is an improved pRM promoter from λ phage (OR-3 is eliminated).
Phage λ is a virus that infects E.coli. After infecting the host E.coli cells, it enters two different life cycles: lytic cycle or lysogenic cycle. In λ phage, the dynamic balance of the separation or combination of CI and early transcription promoter pR controls the lysogenic or lytic pathway of phage. PRM is the promoter of CI, which maintains inhibitor, and only transcribes CI gene. The wild pRM promoter contains OR1, OR2 and OR3 sites, so it can be activated and inhibited by CI protein from the same phage. CI activates the transcription of this promoter at low concentration (binding to OR1 and OR2), but inhibits the transcription at high concentration (binding to OR3).
The improved promoter pRM' can be activated by Protein CI, but not inhibited by Protein CI.
In addition, ssrA-LVA tag was added to the downstream of mRFP1 and LacI in order to allow the protein to be degraded in time.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1032
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1032
Illegal NotI site found at 1931 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1032
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1032
Illegal NgoMIV site found at 548
Illegal AgeI site found at 125
Illegal AgeI site found at 1813 - 1000COMPATIBLE WITH RFC[1000]
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