Part:BBa_K3982000

6xHis - Tagged Cas14a1 with TEV site

Cas14a1 is a type V-F exceptionally compact RNA-guided nuclease. Cas14 forms a complex with RNA-guided nuclease and it can cleave ssDNA without PAM specificity. It comes tagged with 6xHis and has the Tobacco Etch Virus (TEV) protease and cleavage site for easy purification. In our project CODE M, Cas14a1 and sgRNA complex is used to detect SNPs in the Mycobacterium tuberculosis genome in a sputum sample. Upon detection, it triggers cleavage of a Fluorescence-Quencher pair ssDNA reporter and thus fluorescence is emitted which confirms the presence of SNPs in the genome.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 2047
  Illegal EcoRI site found at 3487
  Illegal XbaI site found at 2062
  Illegal PstI site found at 14
  Illegal PstI site found at 2277
  Illegal PstI site found at 2443
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 2047
  Illegal EcoRI site found at 3487
  Illegal PstI site found at 14
  Illegal PstI site found at 2277
  Illegal PstI site found at 2443
  Illegal NotI site found at 7
  Illegal NotI site found at 2053
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 2047
  Illegal EcoRI site found at 3487
  Illegal BamHI site found at 3081
  Illegal XhoI site found at 1031
  Illegal XhoI site found at 1923
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 2047
  Illegal EcoRI site found at 3487
  Illegal PstI site found at 14
  Illegal PstI site found at 2277
  Illegal PstI site found at 2443
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 2047
  Illegal EcoRI site found at 3487
  Illegal XbaI site found at 2062
  Illegal PstI site found at 14
  Illegal PstI site found at 2277
  Illegal PstI site found at 2443
  Illegal AgeI site found at 2737
  
• 1000
  COMPATIBLE WITH RFC[1000]