pcDNA3.1-CRISPR-Puro
(Plasmid #208646)

• Purpose: (Empty Backbone) pcDNA3.1 based empty vector for transient transfection of sgRNA-cas9. U6 promoter ~ puro Res cloned from lentiCRISPRv2 (Addgene#52961), lenti virus sequences not included.
• Depositing Lab: Quanfu Ma
• Publication: RBM24 interacts with RPL3 and regulates RPL3-eIF6 interaction (unpublished)

Backbone

• Vector backbone: pcDNA3.1 (-)
• Backbone manufacturer: Thermo Fisher (Invitrogen)
• Vector type: Mammalian Expression, CRISPR
• Promoter: CMV and U6 in tandem
• Selectable markers: Neomycin (select with G418), Puromycin
• Tags / Fusion Proteins:
  • Flag (C terminal on insert)
  • p2A (C terminal on insert)
  • Puromycin resistant gene (C terminal on insert)
  • Clawed frog nucleoplasmin NLS (C terminal on insert)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Growth instructions: We use stbl3
• Copy number: High Copy

Resource Information

• Supplemental Documents:
  • pcDNA3.1-CRISPR-Puro.gb
• A portion of this plasmid was derived from a plasmid made by: The U6p-BsmBIflanking-Cas9-NLS-p2A-PuroRes cassette was cloned from lentiCRISPRv2 (Addgene#52961), lenti virus related sequences was not included.

Terms and Licenses

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

Depositor Comments

Lenti virus related sequences was not included, for use at Biosafety Level 1. Clone sgRNA as discribed in the document of lentiCRISPRv2 with BsmBI. Note that CMV enhancer-promoter in pcDNA3.1 is not removed, thus the sgRNA would be promoted by tandem CMV-U6 promoter.