Protein_Domain

Part:BBa_K2017000:Design

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


C-split Cas9 + DnaE C-intein


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2020
    Illegal PstI site found at 409
    Illegal PstI site found at 643
    Illegal PstI site found at 1855
    Illegal PstI site found at 2159
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2020
    Illegal PstI site found at 409
    Illegal PstI site found at 643
    Illegal PstI site found at 1855
    Illegal PstI site found at 2159
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2020
    Illegal BamHI site found at 203
    Illegal XhoI site found at 1741
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2020
    Illegal PstI site found at 409
    Illegal PstI site found at 643
    Illegal PstI site found at 1855
    Illegal PstI site found at 2159
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2020
    Illegal PstI site found at 409
    Illegal PstI site found at 643
    Illegal PstI site found at 1855
    Illegal PstI site found at 2159
    Illegal NgoMIV site found at 937
  • 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 C-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.