Lenti-AsCpf1-Blast
(Plasmid #84750)

• Purpose: Expresses human codon-optimized AsCpf1 protein and blasticidin resistance from EFS promoter. Lentiviral backbone.
• Depositing Lab: Hyongbum Kim
• Publication: Kim et al Nat Methods. 2016 Dec 19. doi: 10.1038/nmeth.4104.

Backbone

• Vector backbone: lentiCas9-Blast(Addgene#:52962)
• Backbone size w/o insert (bp): 8700
• Total vector size (bp): 12670
• 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
• Growth instructions: Use SapI digest to check for unwanted recombination of lentiviral plasmid. Only amplify in RecA- bacteria (eg. Stbl3).
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: AsCpf1
• Alt name: Cpf1
• Alt name: AsCas12a
• Species: Acidaminococcus_sp_BV3L6
• Insert Size (bp): 3970
• Promoter: EFS-NS
• Tag / Fusion Protein:
  • NLS-3xHA (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AgeI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: pLLU-F (5'-gggacagcagagatccagtt-3')
• 3′ sequencing primer: blast-R (5'-gctctttcaatgagggtgga-3'​)

Resource Information

• Supplemental Documents:
  • Lenti-AsCpf1-Blast.gb
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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