pCSN071
(Plasmid #166729)

• Purpose: Single copy yeast plasmid expressing DNase-dead dLbCas12a E925A fused to Mxi1 and NLS, codon optimized for Saccharomyces cerevisiae
• Depositing Lab: Rene Verwaal
• Publication: Ciurkot et al Nucleic Acids Res. 2021 Jul 1. pii: 6312738. doi: 10.1093/nar/gkab529.

Backbone

• Vector backbone: pRS414
• Backbone size w/o insert (bp): 7206
• Total vector size (bp): 11102
• Vector type: Yeast Expression, CRISPR
• Selectable markers: TRP1 ; KanMX

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: dCas12a-Mxi1-NLS
• Alt name: dCpf1-Mxi1-NLS
• Species: Synthetic; Lachnospiraceae bacterium ND2006
• Insert Size (bp): 3897
• Mutation: E925A
• Promoter: S. cerevisiae TEF1
• Tags / Fusion Proteins:
  • Mxi1 (C terminal on insert)
  • SV40 NLS (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ATGTCTAAGTTGGAAAAATTCACCA
• 3′ sequencing primer: TTATACCTTTCTCTTCTTCTTTGG

Resource Information

• Supplemental Documents:
  • pCSN071.gb

Terms and Licenses

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