pSSA-RPG
(Plasmid #85932)

• Purpose: Constitutively express DsRed and conditionally express Puro and EGFP. Used to test transfection efficiency and enrich genetically modified cells
• Depositing Lab: Zhiying Zhang
• Publication: Ren et al Cell Mol Life Sci. 2015 Jul;72(14):2763-72. doi: 10.1007/s00018-015-1874-6. Epub 2015 Mar 1.

Backbone

• Vector backbone: pXL-Bac II
• Vector type: Mammalian Expression ; dual-reporter surrogate
• 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: pPB-CMV-DsRed-polyA-CAG-SSA. Puro-T2A-EGFP
• Alt name: DsRed, CAG-SSA. Puro-T2A-EGFP

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: pCAG-F (5'-GCAACGTGCTGGTTATTGTG-3')
• 3′ sequencing primer: SV40pA-R

Resource Information

• Supplemental Documents:
  • Schematic drawing for dual surrogate reporter
  • Annotated GenBank file
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

Puromycin and EGFP will not be expressed until the interrupted puro gene is repaired by single strand annealing (SSA) pathway after a nuclease generates double strand breaks (DSBs).

Nuclease target sequence can be annealed and inserted into the backbone between BamHI and NotI sites using restriction enzymes or Gibson cloning methods.