Coding

Part:BBa_K2429024:Design

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


phEF1a L. shahii dCas13a


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1476
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3076
    Illegal PstI site found at 3445
    Illegal PstI site found at 4174
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1476
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3076
    Illegal PstI site found at 3445
    Illegal PstI site found at 4174
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1476
    Illegal BglII site found at 569
    Illegal BglII site found at 1820
    Illegal BglII site found at 2444
    Illegal BglII site found at 2840
    Illegal BglII site found at 3131
    Illegal BglII site found at 3221
    Illegal BglII site found at 4382
    Illegal BamHI site found at 1183
    Illegal BamHI site found at 5464
    Illegal XhoI site found at 968
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1476
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3076
    Illegal PstI site found at 3445
    Illegal PstI site found at 4174
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1476
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3076
    Illegal PstI site found at 3445
    Illegal PstI site found at 4174
    Illegal NgoMIV site found at 703
    Illegal NgoMIV site found at 5421
    Illegal NgoMIV site found at 5440
    Illegal AgeI site found at 81
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1806
    Illegal SapI.rc site found at 1912
    Illegal SapI.rc site found at 2782
    Illegal SapI.rc site found at 3616


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