Generator
Part:BBa_K1179002:Design
Designed by: Brandon Nadres Group: iGEM13_MIT (2013-09-17)
Hef1A_Cas9-VP16
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 4539
Illegal PstI site found at 315
Illegal PstI site found at 820
Illegal PstI site found at 2928
Illegal PstI site found at 3162
Illegal PstI site found at 4374
Illegal PstI site found at 4678 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4539
Illegal PstI site found at 315
Illegal PstI site found at 820
Illegal PstI site found at 2928
Illegal PstI site found at 3162
Illegal PstI site found at 4374
Illegal PstI site found at 4678 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4539
Illegal BglII site found at 569
Illegal BglII site found at 1904
Illegal BamHI site found at 2722
Illegal XhoI site found at 968
Illegal XhoI site found at 4260 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 4539
Illegal PstI site found at 315
Illegal PstI site found at 820
Illegal PstI site found at 2928
Illegal PstI site found at 3162
Illegal PstI site found at 4374
Illegal PstI site found at 4678 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 4539
Illegal PstI site found at 315
Illegal PstI site found at 820
Illegal PstI site found at 2928
Illegal PstI site found at 3162
Illegal PstI site found at 4374
Illegal PstI site found at 4678
Illegal NgoMIV site found at 703
Illegal NgoMIV site found at 3456
Illegal AgeI site found at 81 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
For people interested in using this part, the Cas9-VP16 has only been used as a transactivator on a minimal CMV promoter.
Source
This part expresses a fusion between the mutant DNA-binding Cas9 protein from Streptococcus pyogenes and the viral VP16 transactivation domain from the herpes simplex virus. Was constructed from performing a gateway reaction with a Hef1a promoter and the Cas9-VP16 entry vector.