Composite

Part:BBa_K1795002:Design

Designed by: Andrew D Halleran, Joseph L Maniaci   Group: iGEM15_William_and_Mary   (2015-09-17)
Revision as of 00:25, 18 September 2015 by Registry (Talk | contribs)

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E. Coli Codon Optimized fdCAS9 under R0010


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1016
    Illegal PstI site found at 2642
    Illegal PstI site found at 3884
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1016
    Illegal PstI site found at 2642
    Illegal PstI site found at 3884
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1216
    Illegal BglII site found at 1272
    Illegal BglII site found at 1414
    Illegal BglII site found at 1962
    Illegal BglII site found at 3802
    Illegal BglII site found at 4041
    Illegal BamHI site found at 3604
    Illegal BamHI site found at 3965
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1016
    Illegal PstI site found at 2642
    Illegal PstI site found at 3884
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1016
    Illegal PstI site found at 2642
    Illegal PstI site found at 3884
    Illegal NgoMIV site found at 4186
    Illegal AgeI site found at 3530
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We made sure to not target the sequence that is promoting the gRNA


Source

The basic template for the sgRNA sequence was taken from the supplementary information of Qi LS, Larson MH, Gilbert LA, et al. Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell. 2013;152(5):1173-1183. doi:10.1016/j.cell.2013.02.022.

References