Coding
Part:BBa_K1179051:Design
Designed by: Brandon Nadres Group: iGEM13_MIT (2013-09-15)
VC155-Cas9(D10A and H840A mutant)
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 4137
Illegal EcoRI site found at 4498
Illegal PstI site found at 558
Illegal PstI site found at 2526
Illegal PstI site found at 2760
Illegal PstI site found at 3972
Illegal PstI site found at 4276 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4137
Illegal EcoRI site found at 4498
Illegal PstI site found at 558
Illegal PstI site found at 2526
Illegal PstI site found at 2760
Illegal PstI site found at 3972
Illegal PstI site found at 4276 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4137
Illegal EcoRI site found at 4498
Illegal BglII site found at 565
Illegal BglII site found at 1502
Illegal BamHI site found at 1
Illegal BamHI site found at 2320
Illegal XhoI site found at 3858 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 4137
Illegal EcoRI site found at 4498
Illegal PstI site found at 558
Illegal PstI site found at 2526
Illegal PstI site found at 2760
Illegal PstI site found at 3972
Illegal PstI site found at 4276 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 4137
Illegal EcoRI site found at 4498
Illegal PstI site found at 558
Illegal PstI site found at 2526
Illegal PstI site found at 2760
Illegal PstI site found at 3972
Illegal PstI site found at 4276
Illegal NgoMIV site found at 3054
Illegal AgeI site found at 4470 - 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.