Protein_Domain

Part:BBa_K2017001:Design

Designed by: Monica Victoria Gutierrez Salazar   Group: iGEM16_Valencia_UPV   (2016-10-03)


N-split Cas9 + DnaE N-intein


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 268
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 268
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 275
    Illegal BglII site found at 1212
    Illegal BglII site found at 2192
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 268
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 268
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

As the part needs to be inserted in a viral vector, the overhangs have to be designed accordingly to the viral vector that is going to be used. The synthesis of N-split Cas9 could not be made in one gBlock, as it is too big. Hence, the half protein had to be splitted in two other parts for its synthesis.


Source

The protein domain is synthesized with the intein

References

Truong, Dong-Jiunn Jeffery et al. "Development Of An Intein-Mediated Split–Cas9 System For Gene Therapy". Nucleic Acids Res 43.13 (2015): 6450-6458. Web. 3 Oct. 2016.