Coding

Part:BBa_K4367013:Design

Designed by: Anders Kllberg   Group: iGEM22_Chalmers-Gothenburg   (2022-10-11)


dCas9-cTEV


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 4358
    Illegal BamHI site found at 4687
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 14
    Illegal BsaI.rc site found at 4694
    Illegal SapI.rc site found at 4594


Design Notes

Effort was put in to find a good length of the linker between dCas9 and nTEV. Also, effort was put into finding good binding sites for dCas9 with gRNA, in relation to binding strength, off-target binding, and distance between the two binding sites of dCas9 (The split halves of TEVp must reach each other).


Source

dCas9 is a variant of Cas9 from the CRISPR/Cas9 system. cTEV is a half of split TEVp, which is Tobacco Etch Virus Protease.

References