Coding
PhyB-908-(

Part:BBa_K365011

Designed by: RENAULT Renaud   Group: iGEM10_ESBS-Strasbourg   (2010-09-15)

Phytochrome B (aa 1-908) linked to a trimer of ClpX (aa 61-425)

Phytochrome B (aa 1-908) was fused to a trimer of ClpX in order to tether substrate containing PIF to the protease upon light application, thus promoting its degradation

Background:

This final construct will be the inducible part of the protease system. This permits the linking of the phytochrome B to the Clpx trimer which forms the light-dependent activation part of our degradation system.


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Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 490
    Illegal BglII site found at 3495
    Illegal BglII site found at 4659
    Illegal BglII site found at 5823
    Illegal BamHI site found at 572
    Illegal XhoI site found at 523
    Illegal XhoI site found at 542
    Illegal XhoI site found at 2677
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2062
    Illegal BsaI site found at 2468
    Illegal SapI site found at 739


Conception:

The standards cloning methods were used. This includes digestion of the vector containing linker- ClpX trimer and the PhyB part then the ligation of them and to finish the transformation of competente cells with this plasmid.

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