DNA

Part:BBa_K2429009:Design

Designed by: Nia Myrie   Group: iGEM17_MIT   (2017-10-12)


pENTR L. shahii dCas13a-DDX6


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 5835
    Illegal PstI site found at 1867
    Illegal PstI site found at 2236
    Illegal PstI site found at 2965
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 5835
    Illegal PstI site found at 1867
    Illegal PstI site found at 2236
    Illegal PstI site found at 2965
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 5835
    Illegal BglII site found at 611
    Illegal BglII site found at 1235
    Illegal BglII site found at 1631
    Illegal BglII site found at 1922
    Illegal BglII site found at 2012
    Illegal BglII site found at 3173
    Illegal BglII site found at 3260
    Illegal BamHI site found at 1
    Illegal BamHI site found at 4255
    Illegal BamHI site found at 4357
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 5835
    Illegal PstI site found at 1867
    Illegal PstI site found at 2236
    Illegal PstI site found at 2965
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 5835
    Illegal PstI site found at 1867
    Illegal PstI site found at 2236
    Illegal PstI site found at 2965
    Illegal NgoMIV site found at 4212
    Illegal NgoMIV site found at 4231
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 597
    Illegal SapI.rc site found at 703
    Illegal SapI.rc site found at 1573
    Illegal SapI.rc site found at 2407
    Illegal SapI.rc site found at 5713


Design Notes

This basic part entry vector is flanked by L1 and L2 sites, which are used to denote a gene. Furthermore, the mutation used to deactivate the protein occurs 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. DDX6 comes from the human genome.

References