Part:BBa_K1385000

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

Italic text==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.

In my experimental plasmid, the CcaR/CcaS/ CpcG2 promoter/ tetR/Tet promoter/EYFP are all on the same plasmid. I was not able to clone the entire plasmid since it is a modified version of Tabor's PJT122 and the CcaR/CcaS genes had illegal restriction sites, so I decided only to biobrick the promoter with tetR. CcaR/CcaS genes can be found on the registry, as well as tet promoters driving reporter proteins; in my case I decided to use EYFP.

The light activation system is as follows:

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). It also needs the CcaR and CcaS genes in order to function. Therefore you need a part such as https://parts.igem.org/Part:BBa_K360051 to work.

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.

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]