Lenti‐Cas9‐2A‐Blast
(Plasmid #73310)

• Purpose: Lentiviral vector expressing human codon-optimized S. pyogenes FLAG-tagged Cas9 and blasticidin resistance from EFS promoter
• Depositing Lab: Jason Moffat
• Publication: Hart et al Cell. 2015 Dec 3;163(6):1515-26. doi: 10.1016/j.cell.2015.11.015. Epub 2015 Nov 25.

Backbone

• Vector backbone: lenti CRISPR pXPR_001
• Backbone manufacturer: Feng Zhang
• Backbone size w/o insert (bp): 6990
• Total vector size (bp): 11214
• Modifications to backbone: Cas9-2A‐Blasticidin vector was modified from lentiCRISPR pXPR_001 (Addgene 49535) by removing the gRNA expressing sequence through NdeI, EcoRI digest and blunt end ligation using large Klenow fragments (NEB) to ligate sticky ends. 2A-­puro was replaced with an amplified 2A-blasticidin sequence with NheI and MluI sites
• Vector type: Mammalian Expression, Lentiviral, CRISPR
• Selectable markers: Blasticidin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Cas9
• Alt name: S. pyogenes CRISPR-Cas9
• Species: Synthetic
• Insert Size (bp): 4224
• Promoter: EFS-NS
• Tag / Fusion Protein:
  • FLAG (N terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: pLLU-F
• 3′ sequencing primer: blast-R

Resource Information

• A portion of this plasmid was derived from a plasmid made by: lentiCRISPR pXPR_001 (Addgene 49535)
• Articles Citing this Plasmid:
  • 43 References

Terms and Licenses

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