Part:BBa_K3602014

Cas12a w. tags, optimized for E. coli K12

TThis part includes the sequence for the cas12a protein with the 6x His Tag/Twin Strep/SUMO tags. The sequence is codon-optimized for expression in E. coli K12. The tags can be used for his-tag purification of the protein after expression from e.g. E. coli TOP10.

Cas12a is a protein with endonuclease activity. In complex with a sgRNA, Cas12a cleaves nearby ssDNA upon complementary binding of a target sequence matching the sgRNA. This can be utilized for detection of different targets, as a DNA reporter can be used for the cleavage. It is noteworthy that IDT was not allowed to produce a part of the Cas12a sequence as it comes from a pathogenic bacterium and therefore requires a special license to ship outside of the United States.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2231
  Illegal BamHI site found at 1476
  Illegal XhoI site found at 1111
  Illegal XhoI site found at 1513
  Illegal XhoI site found at 2974
  Illegal XhoI site found at 3292
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 805
  Illegal AgeI site found at 2296
  Illegal AgeI site found at 3694
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 868