pCAG-T3-eSpCAS9-NG-pA
(Plasmid #131466)

• Purpose: Expresses eSpCAS9-NG nuclease in mammalian cells. Used to modify genome of mouse embroys
• Depositing Lab: Wataru Fujii
• Publication: Fujii et al Sci Rep. 2019 Sep 9;9(1):12878. doi: 10.1038/s41598-019-49394-5.

Backbone

• Vector backbone: pCAGGS
• Total vector size (bp): 9293
• Vector type: Mammalian Expression, CRISPR

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Codon optimized eSpCas9-NG
• Species: Synthetic
• Insert Size (bp): 4218
• Tags / Fusion Proteins:
  • Flag (N terminal on insert)
  • NLS (N terminal on insert)
  • NLS (C terminal on insert)
  • polyA (C terminal on insert)

Terms and Licenses

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

Depositor Comments

This plasmid vector was constructed by mutation PCR of pCAG-T3-hCAS9-pA plasmid vector (Addgene plasmid 48625) according to the amino acid sequences of SpCas9-NG shown in previous study (Nishimasu H et al., Engineered CRISPR-Cas9 nuclease with expanded targeting space. Science, 2018) with high-fidelity eSp variant shown in previous study (Slaymaker IM et al, Rationally engineered Cas9 nucleases with improved specificity. Science, 2016).