RNA

Part:BBa_K3286041

Designed by: Sebastiaan Kuiper   Group: iGEM19_Wageningen_UR   (2019-10-14)
Revision as of 08:47, 17 October 2019 by SJK (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


F. novicida Cpf1 (Cas12a) CRISPR array

Francisella novicida Cpf1 (Cas12a) CRISPR array .

The Cpf1 (Cas12a) CRISPR array of F. novicida, with two repeats and a scrambled (non-targeting) spacer.

New spacers can be generated by BbsI digestion of the CRISPR array and ligation of two annealed oligo's (with 5'-AGAT... and 5'-AGAC overhangs). By doing so, we can generate very fast, cheap and reliable a library of CRISPR arrays*.

(*We have observed that insertion of two spacers (with a repeat in the middle) is also possible with the use of ultramers.)

  1. Zetsche, B., Gootenberg, J. S., Abudayyeh, O. O., Slaymaker, I. M., Makarova, K. S., Essletzbichler, P., … Zhang, F. (2015). Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell, 163(3), 759–771. https://doi.org/10.1016/j.cell.2015.09.038

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None