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:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 4694
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 4719
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1730
Illegal BglII site found at 3410 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 4694
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 4694
Illegal AgeI site found at 55 - 1000INCOMPATIBLE 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.