pRL-TK 4x mut I
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||11322||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4045
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert name4 bulged binding sites for CXCR4 siRNA antisense
MutationMutation I (see paper) on inner 2 binding sites.
/ Fusion Protein
- Rr-luc (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site XbaI (not destroyed)
- 3′ cloning site ApaI (not destroyed)
- 5′ sequencing primer See map (Common Sequencing Primers)
Terms and Licenses
Two identical mutant binding sites, separated by 4 nt, were flanked by two of the orginal bulged binding sites, each 11 nt away.
Two original CXCR4 sites, with XhoI and SpeI restriction sites between them, were inserted into the XbalI site in the 3' UTR of the pRL-TK plasmid. The mutant binding sites were then inserted by ligating annealed oligos into the XhoI and SpeI sites.
See Addgene plasmid 11307 for cloning of binding sites into the backbone, used in Doench, Sharp 2003 paper.
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:pRL-TK 4x mut I was a gift from Phil Sharp (Addgene plasmid # 11322 ; http://n2t.net/addgene:11322 ; RRID:Addgene_11322)
For your References section:Specificity of microRNA target selection in translational repression. Doench JG, Sharp PA. Genes Dev. 2004 Mar 1. 18(5):504-11. 10.1101/gad.1184404 PubMed 15014042