Difference between revisions of "Part:BBa K2127002:Design"

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===Design Notes===
 
===Design Notes===
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Double Stop Codon.
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The His-Tag was developed to isolate the Photosystem II complex and allow for no reduction in activity as measured by electron evolution. A double stop codon and a double terminator were introduced by our team to further ensure termination of transcription in vitro. To allow for easier ability for teams to work and analyze this subunit and its properties, the construct was designed to include an inducible lac promoter under LacI regulation.  
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ORF Check
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When working with this BioBrick it is necessary for teams to ensure there replication host or vector contains a suitable lacI repressor gene.
  
 
===Source===
 
===Source===

Revision as of 13:54, 21 October 2016


CP47 His-tagged with Lac Promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 473
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 597
    Illegal AgeI site found at 1194
    Illegal AgeI site found at 1398
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 918
    Illegal SapI site found at 1595


Design Notes

The His-Tag was developed to isolate the Photosystem II complex and allow for no reduction in activity as measured by electron evolution. A double stop codon and a double terminator were introduced by our team to further ensure termination of transcription in vitro. To allow for easier ability for teams to work and analyze this subunit and its properties, the construct was designed to include an inducible lac promoter under LacI regulation.

When working with this BioBrick it is necessary for teams to ensure there replication host or vector contains a suitable lacI repressor gene.

Source

The psbB gene codes for the CP-47 subunit of the Cyanobacterial Photosystem II in Synechocystis sp. PCC 6803. Using site-directed Mutagenesis Dr Frankel’s team developed a Synechocystis sp. PCC 6803 mutant containing a histidine tag at the C-Terminus of CP47 subunit designated mutant HT-3.

NCBI Reference Sequence: NC_000911.1

References