Part:BBa_K1179028:Design

Cas9-VC155 (D10A and H840 mutant)

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 3867
  Illegal EcoRI site found at 4478
  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
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 3867
  Illegal EcoRI site found at 4478
  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
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 3867
  Illegal EcoRI site found at 4478
  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
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 3867
  Illegal EcoRI site found at 4478
  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
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 3867
  Illegal EcoRI site found at 4478
  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 NgoMIV site found at 2784
  
• 1000
  COMPATIBLE 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 VC155 gBlock and the Cas9 mutant DNA binding entry vector.