Coding
Part:BBa_K1179029:Design
Designed by: Lauren St. Hilaire Group: iGEM13_MIT (2013-09-15)
Cas9-VN155 (D10A and H840A mutant)
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 3867
Illegal EcoRI site found at 4694
Illegal PstI site found at 288
Illegal PstI site found at 2256
Illegal PstI site found at 2490
Illegal PstI site found at 3702
Illegal PstI site found at 4006
Illegal PstI site found at 4413 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 3867
Illegal EcoRI site found at 4694
Illegal PstI site found at 288
Illegal PstI site found at 2256
Illegal PstI site found at 2490
Illegal PstI site found at 3702
Illegal PstI site found at 4006
Illegal PstI site found at 4413 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 3867
Illegal EcoRI site found at 4694
Illegal BglII site found at 295
Illegal BglII site found at 1232
Illegal BamHI site found at 1
Illegal BamHI site found at 2050
Illegal XhoI site found at 3588 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 3867
Illegal EcoRI site found at 4694
Illegal PstI site found at 288
Illegal PstI site found at 2256
Illegal PstI site found at 2490
Illegal PstI site found at 3702
Illegal PstI site found at 4006
Illegal PstI site found at 4413 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 3867
Illegal EcoRI site found at 4694
Illegal PstI site found at 288
Illegal PstI site found at 2256
Illegal PstI site found at 2490
Illegal PstI site found at 3702
Illegal PstI site found at 4006
Illegal PstI site found at 4413
Illegal NgoMIV site found at 2784 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Not applicable
Source
The mutant DNA-binding Cas9 originates from the type II prokaryote CRISPR system in Streptococcus pyogenes. The split venus fluorescent protein is a genetic mutant of the GFP from Aequorea victoria. Constructed by performing a BsaI digestion of an IDT synthesized VN155 gBlock and the Cas9 mutant DNA binding entry vector.