Coding

Part:BBa_K1965050

Designed by: Katja Leben   Group: iGEM16_Slovenia   (2016-10-17)
Revision as of 18:00, 18 October 2016 by NinaJerala (Talk | contribs)

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P5:GS6:PPVs:GS6:cLuc:HA

This construct consists of a C-terminal part of the split luciferase that is fused to the P5 coiled coil via a linker which contains the cleavage substrate for PPV protease. The construct also contains the HA tag on the C-terminus. The P5 coiled coil dimerizes with the AP6 and P6 coiled coils. If its partner coiled coil (AP6 or P6) contains the N-terminal part of the split luciferase, then dimerization of the two coils enables luciferase reconstitution due to the close proximity of the split luciferase parts. However, because the linker contains PPV protease cleavage substrate, the cLuc can be cleaved off in the presence of the PPV protease, preventing reconstitution of the split luciferase.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 139
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None