pPB9P3
(Plasmid #210034)

• Purpose: Contains cloning site for PE3/3b prime editing gRNA constructs, inducible prime editor 2, rtTA-T2A-puro
• Depositing Lab: Michael Talkowski
• Publication: Nuttle et al Cell Rep Methods. 2023 Dec 5:100672. doi: 10.1016/j.crmeth.2023.100672.

Backbone

• Vector backbone: pPB
• Vector type: Mammalian Expression, CRISPR
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Growth instructions: recommend using LB with 100 µg/mL carbenicillin
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: prime editor 2
• Species: Synthetic; Cas9 is from S. pyogenes; M-MLV RT is from the Moloney murine leukemia virus
• Insert Size (bp): 6273
• Promoter: TRE3G
• Tags / Fusion Proteins:
  • SV40 NLS (N terminal on backbone)
  • SV40 NLS (C terminal on backbone)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: N/A

Gene/Insert 2

• Gene/Insert name: rtta-T2A-puro
• Species: Synthetic
• Insert Size (bp): 1395
• Promoter: EF-1α

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: N/A

Resource Information

• Supplemental Documents:
  • pPB9P3.gbk

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • piggyBac Limited Use Label License
  • Tet Systems Limited Use Label License
• Industry Terms:
  • Not Available to Industry