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Inverter

Part:BBa_K1385000

Designed by: Benjamin Huang   Group: iGEM14_WashU_StLouis   (2014-10-07)
Revision as of 20:15, 13 October 2014 by Benjaminhuang (Talk | contribs)

CPCG2 promoter -> TetR

This part incorporates the CpcG2 promoter from Jeff Tabor's 2010 paper "Multichromatic Control of Gene Expression in Escherichia coli" and makes it an inverter. This light regulated promoter expresses TetR in the presence of light. This part should be used in conjunction with https://parts.igem.org/Part:BBa_K1017726 to activate the light regulated plasmid, and a reporter plasmid such as https://parts.igem.org/Part:BBa_J70311 which is a pTet promoter a reporter protein EYFP

Usage and Biology

PcpcG2 is a promoter from the genome of Synechocystis" sp. PCC6803. The promoter comes from Jeffrey Tabor's plasmid pJT122 plasmid, contains the entire region upstream of cpcG2 and downstream of ccaR (Tabor et al. 2011). PcpcG2 is regulated by the light-activation of ccaS/ccaR. The gene downstream to this promoter is transcribed upon activation by ccaS/ccaR, via green light.

The light activation system is as follows:

File:WashU CcaR CcaS.png

This promoter needs to be used in conjunction with the Phycocyanobilin (PCB) which converts Heme and PcyA and Ho1 into the chromophore (why you need to use it with https://parts.igem.org/Part:BBa_K1017726).

When activated by light, this promoter transcribes an output gene, in this case TetR, creating an inverter mechanism for a gene driven by pTet. In our experimental plasmids we had this in conjunction with a pTet promoter driving a reporter protein.

File:WashU PBJ Plasmid.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 279
    Illegal NheI site found at 302
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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