Part:BBa_K2429010:Design

pENTR L. shahii NL-Cas13a-NL

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 4425
  Illegal PstI site found at 1903
  Illegal PstI site found at 2272
  Illegal PstI site found at 3001
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 4425
  Illegal PstI site found at 1903
  Illegal PstI site found at 2272
  Illegal PstI site found at 3001
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 4425
  Illegal BglII site found at 647
  Illegal BglII site found at 1271
  Illegal BglII site found at 1667
  Illegal BglII site found at 1958
  Illegal BglII site found at 2048
  Illegal BglII site found at 3209
  Illegal BglII site found at 3296
  Illegal BamHI site found at 1
  Illegal BamHI site found at 70
  Illegal BamHI site found at 4291
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 4425
  Illegal PstI site found at 1903
  Illegal PstI site found at 2272
  Illegal PstI site found at 3001
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 4425
  Illegal PstI site found at 1903
  Illegal PstI site found at 2272
  Illegal PstI site found at 3001
  Illegal NgoMIV site found at 4248
  Illegal NgoMIV site found at 4267
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 633
  Illegal SapI.rc site found at 739
  Illegal SapI.rc site found at 1609
  Illegal SapI.rc site found at 2443
  

Design Notes

This basic part entry vector is flanked by L1 and L2 sites, which are used to denote a gene. There are glycine-serine link between the dCas13a protein and the nuclear localization sequences.

Source

Sequence for L. shahii Cas13a was found from Addgene(https://www.addgene.org/79150/), which is also where our team received the original plasmid from. See reference for nuclear localization amino acid sequence.

References

[1] Lange, et al. "Classical Nuclear Localization Signals: Definition, Function, and Interaction with Importin α*"