Part:BBa_K1642011
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.
Based on Pdark and HR, we constructed an improved biodesalination system which depends only on the switch between white light and darkness. In the growth stage and working stage, we provide white light, while in the induced expression stage the light source is removed. Darkness leads to starvation and the starvation can inhibit active export of sodium, which is essential for biodesalination in the working stage.(described l in section transport module). The biodesalination process controlled by Pdark is shown in Figure 1.4.3.
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]
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