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:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 4539
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 4564
- 21INCOMPATIBLE 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 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 4539
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 4539
Illegal AgeI site found at 55 - 1000INCOMPATIBLE 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.