Coding

Part:BBa_K2429040:Design

Designed by: Nia Myrie   Group: iGEM17_MIT   (2017-10-24)


pTRE L. shahii dCas13a


Assembly Compatibility:
  • 10
    INCOMPATIBLE 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
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE 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
  • 23
    INCOMPATIBLE 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
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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.


References