Lenti-TurboCas-NLS-Blast
(Plasmid #232346)

• Purpose: Lentiviral expression vector for dCas9 fused to miniTurboNLS
• Depositing Lab: Ali Shilatifard
• Publication: Cenik et al Mol Cell. 2024 Dec 19;84(24):4929-4944.e8. doi: 10.1016/j.molcel.2024.11.007.

Backbone

• Vector backbone: lentiCas9-Blast
• Backbone manufacturer: Feng Zhang
• Backbone size w/o insert (bp): 8149
• Total vector size (bp): 13769
• Vector type: Mammalian Expression, Lentiviral, CRISPR
• Selectable markers: Blasticidin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Growth instructions: Propagate using low-recombination strains, we recommend Stbl3 or NEBStable.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: dCas9
• Alt name: Cas9m4
• Species: Synthetic
• Insert Size (bp): 4875
• Promoter: EF1a
• Tags / Fusion Proteins:
  • 3xHA, miniTurbo, V5 (N terminal on insert)
  • 3xSV40 NLS (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: pLLU-F, tgggcaataccgatcgcca

Resource Information

• Supplemental Documents:
  • Lenti-TurboCas-NLS-Blast.gb

Terms and Licenses

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

Depositor Comments

Cloned by removing insert from LentiCas9-blast from Feng Zheng (Addgene #52962) and replacing with TurboCas insert by Gibson assembly. Addgene observed S522P substitution in Cas9, which is not of concern to plasmid function.