Composite

Part:BBa_K2926086:Design

Designed by: Johanna Opgenoorth   Group: iGEM19_Bielefeld-CeBiTec   (2019-10-21)


Basic Insert to express Cas13a (Lwa) and a short guideRNA array in S. cerevisiae


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 4694
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 4719
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1730
    Illegal BglII site found at 3410
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 4694
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 4694
    Illegal AgeI site found at 55
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2809


Design Notes

Due to the repetitivity of the guide RNA cluster we assembled it using Golden Gate Assembly.


Source

Promoters and terminators were obtained via gene synthesis while Cas13a was excised from Munich 2017s plasmid and the guide RNA cluster was cloned via Golden Gate Assembly.


References