sgOpti-tevoPQ1
(Plasmid #255532)

• Purpose: lenti virus vector expressing a prime editing guide RNA under a human U6 promoter. Contains a truncated evoPQ1 sequence at the 3' end of the pegRNA which was shown to improve editing efficiency.
• Depositing Lab: Jesse Engreitz
• Publication: Rewriting regulatory DNA to dissect and reprogram gene expression. (unpublished)

Backbone

• Vector backbone: sgOpti
• Total vector size (bp): 9025
• Vector type: Lentiviral, CRISPR
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Growth instructions: Use NEB Stable cells. Grow at 30°C. Lentiviral/guide RNA vectors can recombine, so avoid overgrowth and extended culture. Start from a glycerol stock when possible and use fresh transformants.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: prime editing guideRNA
• Alt name: pegRNA
• gRNA/shRNA sequence: (no spacer inserted)
• Promoter: U6 (human U6 promoter)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsmBI (Esp3I) (destroyed during cloning)
• 3′ cloning site: BsmBI (Esp3I) (destroyed during cloning)
• 5′ sequencing primer: U6 promoter primer
• 3′ sequencing primer: sgPuro_RevOuterSeq

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Constructed in-house using sgOpti backbone for prime editing guide RNA cloning.

Terms and Licenses

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