pSicoR-Ef1a-mCh-Puro
(Plasmid #31845)

• Purpose: (Empty Backbone)
• Depositing Lab: Bruce Conklin
• Publication: Salomonis et al Proc Natl Acad Sci U S A. 2010 Jun 8;107(23):10514-9. Epub 2010 May 24.

Backbone

• Vector backbone: pSicoR-mCH
• Backbone manufacturer: Addgene plasmid #21907
• Backbone size (bp): 8141
• Modifications to backbone: The pSicoR-Ef1α-mCh-Puro lentiviral construct was created by replacing the CMV promoter of pSicoR-mCherry (Addgene vector 21907) with Ef1α and adding an in-frame T2A-puromycin-resistance gene cassette following mCherry. A single RNA molecule is generated for mCherry-T2A-puromycin and two polypeptides are produced by the T2A ribosomal skip motif. The shRNA coding oligos are cloned into the HpaI and XhoI restriction sites as described for pSicoR.
• Vector type: Mammalian Expression, Lentiviral, RNAi, Cre/Lox
• Promoter: U6 for shRNA; Ef1alpha for mCherry-puromycin
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Resource Information

• Supplemental Documents:
  • pSicoR-Ef1a-mCh-Puro_Seq
  • pSicoR-Ef1a-mCh-Puro_txt
  • pSicoR-Ef1a-mCh-Puro.gb
• Addgene Notes:
  • Addgene diagnostic digest
• Articles Citing this Plasmid:
  • 19 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The pSicoR-Ef1α-mCh-Puro lentiviral construct was created by replacing the CMV promoter of pSicoR-mCherry (Addgene vector 21907) with Ef1α and adding an in-frame T2A-puromycin-resistance gene cassette following mCherry. A single RNA molecule is generated for mCherry-T2A-puromycin and two polypeptides are produced by the T2A ribosomal skip motif. The shRNA coding oligos are cloned into the HpaI and XhoI restriction sites as described for pSicoR.