Part:BBa_K2340000:Design

dCAS13a linked to GFP with NLS

Assembly Compatibility:

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

  Illegal BglII site found at 1112
  Illegal BglII site found at 2048
  Illegal BglII site found at 2312
  Illegal BglII site found at 2816
  Illegal BglII site found at 3302
  Illegal BglII site found at 3410
  Illegal BglII site found at 3740
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 662
  Illegal SapI.rc site found at 3421
  

Design Notes

Addition of LINK and NLS sequence

Source

The sequence of this biobrick is synthesized by Integrated DNA Technologies (IDT). The p2CT-His-MBP-Lbu_C2c2_R472A_H477A_R1048A_H1053A gene sequence was a gift from Jennifer Doudna (Addgene plasmid # 83485¹). Three nucleotides were edited to remove the illegal restriction enzyme cutting sites. To ensure efficient protein expression a codon usage table was used². The sequence of the GFP gene was obtained from the biobrick library (BBa_K648013).

References

¹Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. East-Seletsky A, O'Connell MR, Knight SC, Burstein D, Cate JH, Tjian R, Doudna JA. Nature. 2016 Sep 26. doi: 10.1038/nature19802. 10.1038/nature19802 PubMed 27669025

²GenScript (2017). GenScript Codon Usage Frequence Table Tool. [WWW document]. URL https://www.genscript.com/tools/codon-frequency-table