DNA
dC2c2

Part:BBa_K2429007:Design

Designed by: Nia Myrie   Group: iGEM17_MIT   (2017-10-12)
Revision as of 14:52, 26 October 2017 by Nmyrie (Talk | contribs) (→‎Design Notes)

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pENTR L. shahii dCas13a


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 294
    Illegal PstI site found at 1894
    Illegal PstI site found at 2263
    Illegal PstI site found at 2992
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 294
    Illegal PstI site found at 1894
    Illegal PstI site found at 2263
    Illegal PstI site found at 2992
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 294
    Illegal BglII site found at 638
    Illegal BglII site found at 1262
    Illegal BglII site found at 1658
    Illegal BglII site found at 1949
    Illegal BglII site found at 2039
    Illegal BglII site found at 3200
    Illegal BglII site found at 3287
    Illegal BamHI site found at 1
    Illegal BamHI site found at 4282
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 294
    Illegal PstI site found at 1894
    Illegal PstI site found at 2263
    Illegal PstI site found at 2992
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 294
    Illegal PstI site found at 1894
    Illegal PstI site found at 2263
    Illegal PstI site found at 2992
    Illegal NgoMIV site found at 4239
    Illegal NgoMIV site found at 4258
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 624
    Illegal SapI.rc site found at 730
    Illegal SapI.rc site found at 1600
    Illegal SapI.rc site found at 2434


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.


References