Plasmid 31845: pSicoR-Ef1a-mCh-Puro

• Gene/insert name: None
• Vector backbone: pSicoR-mCH
  (Search Vector Database)
• Backbone manufacturer: Addgene plasmid #21907
• Vector type: Mammalian Expression, Lentiviral, RNAi, Cre/Lox
• 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.
• Promoter: U6 for shRNA; Ef1alpha for mCherry-puromycin
• Bacterial resistance(s) Ampicillin
• Growth strain(s) Stbl3
• Growth temperature (℃): 37
• High or low copy: High Copy
• Selectable markers: Puromycin
• Sequence: View sequences (2)
• Map: View map
• Supplemental document: pSicoR-Ef1a-mCh-Puro_Seq (application/pdf)
• Supplemental document: pSicoR-Ef1a-mCh-Puro_txt (text/plain)
• Supplemental document: pSicoR-Ef1a-mCh-Puro.gb (application/octet-stream)
• Principal Investigator: Bruce Conklin
• Terms and Licenses: MTA
  Clontech Limited Use Label License

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.

Article: Alternative splicing regulates mouse embryonic stem cell pluripotency and differentiation. Salomonis et al (Proc Natl Acad Sci U S A. 2010 Jun 8;107(23):10514-9. Epub 2010 May 24. PubMed)