Generator

Part:BBa_K1179002:Design

Designed by: Brandon Nadres   Group: iGEM13_MIT   (2013-09-17)


Hef1A_Cas9-VP16


Assembly Compatibility:
  • 10
    INCOMPATIBLE 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
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE 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
  • 23
    INCOMPATIBLE 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
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE 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.

References