Difference between revisions of "Part:BBa K1385000:Design"
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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 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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 279
Illegal NheI site found at 302 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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