Coding
Part:BBa_K2429040:Design
Designed by: Nia Myrie Group: iGEM17_MIT (2017-10-24)
pTRE L. shahii dCas13a
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 6
Illegal EcoRI site found at 322
Illegal EcoRI site found at 632
Illegal XbaI site found at 30
Illegal PstI site found at 2232
Illegal PstI site found at 2601
Illegal PstI site found at 3330 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 6
Illegal EcoRI site found at 322
Illegal EcoRI site found at 632
Illegal PstI site found at 2232
Illegal PstI site found at 2601
Illegal PstI site found at 3330 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 6
Illegal EcoRI site found at 322
Illegal EcoRI site found at 632
Illegal BglII site found at 976
Illegal BglII site found at 1600
Illegal BglII site found at 1996
Illegal BglII site found at 2287
Illegal BglII site found at 2377
Illegal BglII site found at 3538
Illegal BglII site found at 3625
Illegal BamHI site found at 339
Illegal BamHI site found at 4620 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 6
Illegal EcoRI site found at 322
Illegal EcoRI site found at 632
Illegal XbaI site found at 30
Illegal PstI site found at 2232
Illegal PstI site found at 2601
Illegal PstI site found at 3330 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 6
Illegal EcoRI site found at 322
Illegal EcoRI site found at 632
Illegal XbaI site found at 30
Illegal PstI site found at 2232
Illegal PstI site found at 2601
Illegal PstI site found at 3330
Illegal NgoMIV site found at 4577
Illegal NgoMIV site found at 4596 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 962
Illegal SapI.rc site found at 1068
Illegal SapI.rc site found at 1938
Illegal SapI.rc site found at 2772
Design Notes
This part is human codon optimized. Furthermore, the mutation used to deactivate the protein occurs (starting from the beginning of the dCas13a gene) from base pairs 3830 to 3832, in which the bases were changed from CGG to GCC.
Source
The catalytically active sequence of the protein came from L. shahii Cas13a on Addgene(https://www.addgene.org/79150/), which is also where our team received the original plasmid from.