pPC1000
(Plasmid #174829)

• Purpose: Lentiviral Repair-seq vector containing CRISPRi sgRNA and SaPE2 prime edit site
• Depositing Lab: David Liu
• Publication: Chen et al Cell. 2021 Oct 28;184(22):5635-5652.e29. doi: 10.1016/j.cell.2021.09.018.

Backbone

• Vector backbone: Lentiviral
• Vector type: Mammalian Expression, Lentiviral, CRISPR
• Selectable markers: Puromycin ; BFP

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 1

• Gene/Insert name: CRISPRi SpCas9 sgRNA cassette
• Alt name: EGFP-targeting single guide RNA (modified F+E SpCas9 scaffold) for CRISPRi
• Species: Synthetic
• Insert Size (bp): 112
• Mutation: Detailed in manuscript
• Promoter: mouse U6

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: BstXI (not destroyed)
• 3′ cloning site: BlpI (not destroyed)
• 5′ sequencing primer: mouse U6-forward

Gene/Insert 2

• Gene/Insert name: PuroR-P2A-BFP
• Alt name: Puromycin N-acetyl-transferase-P2A-blue fluorescent protein
• Species: Synthetic
• Insert Size (bp): 1368
• Promoter: EF1a

Cloning Information for Gene/Insert 2

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: EF1a-forward
• 3′ sequencing primer: WPRE-reverse

Resource Information

• Supplemental Documents:
  • pPC1000.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry