pET-21a_3xNLS_SpCas9_protein_expression
(Plasmid #114365)

• Purpose: Protein expression plasmid for 3xNLS SpCas9 in pET-21a backbone
• Depositing Lab: Scot Wolfe
• Publication: Wu et al Nat Med. 2019 May;25(5):776-783. doi: 10.1038/s41591-019-0401-y. Epub 2019 Mar 25.

Backbone

• Vector backbone: pET-21 a
• Backbone size w/o insert (bp): 5400
• Total vector size (bp): 9705
• 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: 3xNLS_SpCas9
• Species: Synthetic; S. pyogenes
• Insert Size (bp): 4262
• Promoter: T7
• Tags / Fusion Proteins:
  • NLS (N terminal on insert)
  • NLS (C terminal on insert)
  • NLS (C terminal on insert)

Cloning Information

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

Resource Information

• Supplemental Documents:
  • 3nls-sp
• Articles Citing this Plasmid:
  • 8 References

Terms and Licenses

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

Depositor Comments

This plasmid is also associated with the following publication:
PMID 30911135: Highly efficient therapeutic gene editing of human hematopoietic stem cells