pNOC_dfnCas12a-KRAB_crRNA NC
(Plasmid #176262)

• Purpose: pNOC episomal plasmid harboring the dead version of humanized fnCas12a gene sequence tagged to the KRAB domain and Nlux and spacer sequence is replaced by the type IIS restriction site for endonucleas
• Depositing Lab: John van der Oost
• Publication: Naduthodi et al ACS Synth Biol. 2021 Dec 17;10(12):3369-3378. doi: 10.1021/acssynbio.1c00329. Epub 2021 Nov 18.

Backbone

• Vector backbone: pUC19
• Vector type: CRISPR, Luciferase, Synthetic Biology ; Expression in microalgae Nannochloropsis oceanica
• Selectable markers: Blasticidin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: humanized fnCas12a
• Species: Francisella novicida
• Mutation: D917A and E1006A mutations to inactivate the endonuclease activity of Cas12a
• Tags / Fusion Proteins:
  • luciferase (C terminal on insert)
  • Kruppel associated box domain (C terminal on insert)

Resource Information

• Supplemental Documents:
  • pnoc_dfncas12a-krab_crrna-nc.gb
• A portion of this plasmid was derived from a plasmid made by: The backbone of plasmid was a gift from Eva Farre from Michigan State University. And the Cas12a sequence was obtained from the Addgene plasmid # 69976

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry