Difference between revisions of "Part:BBa K1642011"

 
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<partinfo>BBa_K1642011 short</partinfo>
 
<partinfo>BBa_K1642011 short</partinfo>
  
Pdark-HR
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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.
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Up and Down are sequences uesd for homologous recombination.
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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.
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RBS is the RNA binding site, which will hopefully increase the expression level of HR.
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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.
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Terminator is the transcription terminator for the E.coli RNA polymerase.
  
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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.
 
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===Usage and Biology===
 
===Usage and Biology===

Revision as of 11:46, 16 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 uesd 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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 611
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3215
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE WITH RFC[1000]