Coding

Part:BBa_K2429028:Design

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


phEF1a L. shahii NL dCas13a


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 5604
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3085
    Illegal PstI site found at 3454
    Illegal PstI site found at 4183
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 5604
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3085
    Illegal PstI site found at 3454
    Illegal PstI site found at 4183
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 5604
    Illegal BglII site found at 569
    Illegal BglII site found at 1829
    Illegal BglII site found at 2453
    Illegal BglII site found at 2849
    Illegal BglII site found at 3140
    Illegal BglII site found at 3230
    Illegal BglII site found at 4391
    Illegal BamHI site found at 1183
    Illegal BamHI site found at 1252
    Illegal BamHI site found at 5473
    Illegal XhoI site found at 968
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 5604
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3085
    Illegal PstI site found at 3454
    Illegal PstI site found at 4183
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 5604
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3085
    Illegal PstI site found at 3454
    Illegal PstI site found at 4183
    Illegal NgoMIV site found at 703
    Illegal NgoMIV site found at 5430
    Illegal NgoMIV site found at 5449
    Illegal AgeI site found at 81
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1815
    Illegal SapI.rc site found at 1921
    Illegal SapI.rc site found at 2791
    Illegal SapI.rc site found at 3625


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. We also put in glycine-serine linker sequences to connect the NLSs

References