pPB-US-ECasE
(Plasmid #83961)

• Purpose: Piggyac transposon containing tamoxifen inducible Cas9 and a gRNA cassette.
• Depositing Lab: Eleanor Chen
• Publication: Chen lab CRISPR plasmids (unpublished)

Backbone

• Vector backbone: Piggybac Transposon
• Backbone manufacturer: Sytem Biosciences
• Vector type: Mammalian Expression
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: ERT2-spCas9-ERT2
• Insert Size (bp): 7970
• Mutation: please see depositor comments below
• Promoter: U6 (gRNA), EF1a (ERT2-Cas9-ERT2)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: unknown

Resource Information

• Supplemental Documents:
  • genbank file
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Please note that mutation R1058W in Cas9 was found during Addgene's quality control. The depositor noted that this mutation does NOT affect plasmid function.

The plasmid expresses a ERT2-SpCas9-ERT2 tamoxifen inducible Cas9 driven by the EF-1a promoter. T2A linked puromucin enables stable selection of the piggybac transposon. CRISPR gRNAS can be cloned into the vector at the BbsI cloning sites.