pEN019-SpCas9-hiNLS-t-M1M4
(Plasmid #234456)

• Purpose: Expresses hiNLS variant t-M1M4 inside 0C triNLS-SpCas9-NGG
• Depositing Lab: Ross Wilson
• Publication: Noel et al CRISPR J. 2025 Apr;8(2):105-119. doi: 10.1089/crispr.2024.0080. Epub 2025 Mar 31.

Backbone

• Vector backbone: pEC-K-MBP
• Backbone manufacturer: pET-based custom vector
• Backbone size w/o insert (bp): 4728
• Total vector size (bp): 9237
• Modifications to backbone: removed HA and MBP
• Vector type: Bacterial Expression, CRISPR

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: SpCas9 hiNLS t-M1M4
• Alt name: SpCas9 0C triNLS hiNLS M1M4
• Species: Streptococcus pyogenes
• Insert Size (bp): 4509
• Mutation: Cys at positions 80 and 574 have been mutated to Ser; Met at position 1 has been mutated to Ser; inserted hiNLS c-Myc (t-M1M4, hairpin internal)
• Tags / Fusion Proteins:
  • His6 (N terminal on backbone)
  • c-Myc (N terminal on insert)
  • SV40 (C terminal on insert)
  • Nucleoplasmin (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: T7 terminator

Resource Information

• Supplemental Documents:
  • pEN019-SpCas9-hiNLS-t-M1M4.gbk

Terms and Licenses

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