|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||31848||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerAddgene plasmid #31845
- Backbone size w/o insert (bp) 8181
Vector typeMammalian Expression, Lentiviral, RNAi, Cre/Lox
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameGFP RNAi
- Promoter U6 for shRNA; Ef1alpha for mCherry-puromycin
- Cloning method Restriction Enzyme
- 5′ cloning site HpaI (destroyed during cloning)
- 3′ cloning site XhoI (not destroyed)
- 5′ sequencing primer U6 (Common Sequencing Primers)
This vector can be used as an shRNA control. 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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pSicoR-Ef1a-mCh-Puro-GFPi was a gift from Bruce Conklin (Addgene plasmid # 31848 ; http://n2t.net/addgene:31848 ; RRID:Addgene_31848)
For your References section:Alternative splicing regulates mouse embryonic stem cell pluripotency and differentiation. Salomonis N, Schlieve CR, Pereira L, Wahlquist C, Colas A, Zambon AC, Vranizan K, Spindler MJ, Pico AR, Cline MS, Clark TA, Williams A, Blume JE, Samal E, Mercola M, Merrill BJ, Conklin BR. Proc Natl Acad Sci U S A. 2010 Jun 8;107(23):10514-9. Epub 2010 May 24. 10.1073/pnas.0912260107 PubMed 20498046