Coding
Part:BBa_K1179057:Design
Designed by: Brandon Nadres Group: iGEM13_MIT (2013-09-15)
VN173-Cas9(D10A and H840A mutant)
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 4401
Illegal EcoRI site found at 4762
Illegal PstI site found at 243
Illegal PstI site found at 822
Illegal PstI site found at 2790
Illegal PstI site found at 3024
Illegal PstI site found at 4236
Illegal PstI site found at 4540 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4401
Illegal EcoRI site found at 4762
Illegal PstI site found at 243
Illegal PstI site found at 822
Illegal PstI site found at 2790
Illegal PstI site found at 3024
Illegal PstI site found at 4236
Illegal PstI site found at 4540 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4401
Illegal EcoRI site found at 4762
Illegal BglII site found at 829
Illegal BglII site found at 1766
Illegal BamHI site found at 1
Illegal BamHI site found at 2584
Illegal XhoI site found at 4122 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 4401
Illegal EcoRI site found at 4762
Illegal PstI site found at 243
Illegal PstI site found at 822
Illegal PstI site found at 2790
Illegal PstI site found at 3024
Illegal PstI site found at 4236
Illegal PstI site found at 4540 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 4401
Illegal EcoRI site found at 4762
Illegal PstI site found at 243
Illegal PstI site found at 822
Illegal PstI site found at 2790
Illegal PstI site found at 3024
Illegal PstI site found at 4236
Illegal PstI site found at 4540
Illegal NgoMIV site found at 3318
Illegal AgeI site found at 4734 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Not applicable
Source
Was constructed by digestion and ligation of BsaI sites on IDT synthesized split venus proteins and the DNA binding mutant Cas9 entry vector with N-terminal BsaI site. The Cas9 comes from the type II prokaryote CRISPR system in Streptococcus pyogenes. The split venus fluorescence protein is a genetic mutant of GFP which comes from the jellyfish Aequorea victoria.