Purpose(Empty Backbone) Mouse genomic ROSA26 targeting vector, expresses EGFP, U6promoter to drive short RNAs, pgkneo, cassette flanked by loxP sites
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||54479||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerPhil Soriano, Addgene Plasmid #21714
Modifications to backboneModified by deleting the HpaI site, converting the XhoI site to AscI, and cloning a splice acceptor-GFP-polyA into the XbaI site. Cre-lox regulated U6 cassette derived from pSICOR, Addgene plasmid 11579, (Ventura et al., 2004) was modified by replacing the lentiviral GFP gene with drug selection markers (pgk-neo), and cloned into the XbaI site 3' of GFP. Unique HpaI and XhoI sites were maintained for single step short hairpin oligonucleotide cloning.
Vector typeMammalian Expression, Mouse Targeting, RNAi, Cre/Lox
- Promoter U6
Selectable markersNeomycin (select with G418)
/ Fusion Proteins
- SA-EGFP-polyA (N terminal on backbone)
- loxP (N terminal on backbone)
- U6 promoter (N terminal on backbone)
- pgk-neo (C terminal on backbone)
- loxP (C terminal on backbone)
Growth in Bacteria
Copy numberHigh Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer RUSH-for CACAAAAGGAAACTCAC CCTAA
- 3′ sequencing primer RUSH-rev CTAAAGCGCATGCTCCA GAC (Common Sequencing Primers)
Terms and Licenses
Unique HpaI and XhoI sites were maintained for single step short hairpin oligonucleotide cloning.
Using the primers described above yields 231bp and 177bp for positive and negative products, respectively.
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:pRUSH was a gift from Laurie Jackson-Grusby (Addgene plasmid # 54479 ; http://n2t.net/addgene:54479 ; RRID:Addgene_54479)
For your References section:RUSH and CRUSH: a rapid and conditional RNA interference method in mice. Brown JR, Zetsche B, Jackson-Grusby L. Genesis. 2014 Jan;52(1):39-48. doi: 10.1002/dvg.22718. Epub 2013 Oct 26. 10.1002/dvg.22718 PubMed 24166816
Map uploaded by the depositor.