Composite
Part:BBa_K1795002:Design
Designed by: Andrew D Halleran, Joseph L Maniaci Group: iGEM15_William_and_Mary (2015-09-17)
E. Coli Codon Optimized fdCAS9 under R0010
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1016
Illegal PstI site found at 2642
Illegal PstI site found at 3884 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1016
Illegal PstI site found at 2642
Illegal PstI site found at 3884 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1016
Illegal PstI site found at 2642
Illegal PstI site found at 3884 - 25INCOMPATIBLE 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 - 1000COMPATIBLE 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.