Part:BBa_K4611010

LwaCas13a

Cas13 orthologs, the single effectors of the Class 2 type VI clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated protein (Cas) systems, are RNA-guided ribonucleases1. The CRISPR RNAs (crRNAs) of this family contain a direct repeat stem loop that interacts with the Cas13 protein to form an RNase-inactive binary complex and a spacer sequence that base pairs with the target RNA2. The resulting Cas13–crRNA–target ternary complex undergoes a large-scale conformational change in which two higher eukaryotes and prokaryotes nucleotide-binding (HEPN) domains move toward each other to form a single catalytic pocket to cleave the target RNA. Intriguingly, this catalytic pocket localized on the outer surface of the target-activated Cas13 complex can non-specifically cleave any surrounding RNA molecules in a characteristic ‘collateral effect’2. This target-triggered collateral activity, originally an immune defense mechanism intended to induce host dormancy and prevent the propagation of invading phages, has been rapidly developed for in vitro detection of nucleic acids3.(Yang J...2023) LwaCas13a is a kind of Cas13 protein from Leptotrichia wadei.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 2830
  Illegal PstI site found at 2851
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 2830
  Illegal PstI site found at 2851
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 2830
  Illegal BglII site found at 1286
  Illegal BglII site found at 1496
  Illegal BglII site found at 1763
  Illegal BglII site found at 2528
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 2830
  Illegal PstI site found at 2851
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 2830
  Illegal PstI site found at 2851
  
• 1000
  COMPATIBLE WITH RFC[1000]