Difference between revisions of "Part:BBa K1642011"
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Pdark-HR is a plasmid used to test whether the protein halorhodopsin under the control of P dark works as we expect. | Pdark-HR is a plasmid used to test whether the protein halorhodopsin under the control of P dark works as we expect. | ||
| − | Up and Down are sequences | + | Up and Down are sequences used for homologous recombination. |
Pdark(BBa_K1026009) is a “dark-sensing” promoter combines PcpcG2 and a constitutive promoter from ''E.coli''. Pdark contains binding site of CcaR in PcpcG2, which overlaps with the binding site of RNA polymerase. Therefore when phosphorylated CcaR binds to it, the binding of RNA polymerase to Pdark will be blocked and transcription can’t be initiated. | Pdark(BBa_K1026009) is a “dark-sensing” promoter combines PcpcG2 and a constitutive promoter from ''E.coli''. Pdark contains binding site of CcaR in PcpcG2, which overlaps with the binding site of RNA polymerase. Therefore when phosphorylated CcaR binds to it, the binding of RNA polymerase to Pdark will be blocked and transcription can’t be initiated. | ||
Revision as of 11:12, 18 September 2015
Expression of Halorhodopsin under the control of Pdrak
Pdark-HR is a plasmid used to test whether the protein halorhodopsin under the control of P dark works as we expect.
Up and Down are sequences used for homologous recombination.
Pdark(BBa_K1026009) is a “dark-sensing” promoter combines PcpcG2 and a constitutive promoter from E.coli. Pdark contains binding site of CcaR in PcpcG2, which overlaps with the binding site of RNA polymerase. Therefore when phosphorylated CcaR binds to it, the binding of RNA polymerase to Pdark will be blocked and transcription can’t be initiated.
RBS is the RNA binding site, which will hopefully increase the expression level of HR.
Halorhodopsin (HR) proteins are light-driven inward-directed chloride pumps from halobacteria. We use this as our biodesalination driver which confers cyanobacteria the ability to absorb chloride to a significant degree.
Terminator is the transcription terminator for the E.coli RNA polymerase.
PcpcB is a constitutive promoter in cyanobacteria, and Kana is gene sequence for kanamycin resistance, so that bacteria which are successfully transferred this plasmid can be screened on the plate.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 611
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 3215
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 481
Illegal NgoMIV site found at 783
Illegal NgoMIV site found at 942
Illegal NgoMIV site found at 1263
Illegal AgeI site found at 1134 - 1000COMPATIBLE WITH RFC[1000]