Difference between revisions of "Part:BBa K1385000:Design"
(→‎Use)
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===Use===
 
===Use===
−
To Use this promoter, you need to co-transform with the Phycocyanobilin Plasmid such as BBa_K1017726. Then you have to grow cultures in either green light (535nm) or broad spectrum light. to produce high levels of TetR.
+
To Use this promoter, you need to co-transform with the Phycocyanobilin Plasmid such as BBa_K1017726. You also need the CcaR/CcaS part such as BBa_K360051. And a Ptet promoter driving your desired gene. Then you have to grow cultures in either green light (535nm) or broad spectrum light to produce high levels of TetR.
  
 
===Design Notes===
 
===Design Notes===

Revision as of 20:54, 13 October 2014

CPCG2 promoter -> TetR


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]

Use

To Use this promoter, you need to co-transform with the Phycocyanobilin Plasmid such as BBa_K1017726. You also need the CcaR/CcaS part such as BBa_K360051. And a Ptet promoter driving your desired gene. Then you have to grow cultures in either green light (535nm) or broad spectrum light to produce high levels of TetR.

Design Notes

The spacer region between TetR and cpcG2 can be modified in order to increase or decrease the RBS strength, as TetR start codon is a ways down from the end of the promoter. The region it is so far is because our primers when assembling the plasmid had troublesome secondary structures closer to the promoter and start codon, so we needed a spacer.


Source

cpcG2 promoter Genes from PJT122 (Tabor lab) TetR from a plasmid Ptet-pp* found available in Moon Lab, at Washington University in St. Louis

References

Yuu Hirose et al ( 2008 )"Cyanobacteriochrome CcaS is the green light receptor that induces the expression of phycobilisome linker protein." Proc Natl Acad Sci U S A. 105: 9528–9533.

Tabor, J. J. et al.(2010), " Multichromatic Control of Gene Expression in Escherichia coli", J. Mol. Biol. , doi:10.1016/j.jmb.2010.10.038