Composite

Part:BBa_K2926084:Design

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


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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 4539
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 4564
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 788
    Illegal BglII site found at 1724
    Illegal BglII site found at 1988
    Illegal BglII site found at 2492
    Illegal BglII site found at 2978
    Illegal BglII site found at 3086
    Illegal BglII site found at 3416
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 4539
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 4539
    Illegal AgeI site found at 55
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 3097


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