Addgene - pRL-TK 4x wt Plasmid Data

Plasmid 11313: pRL-TK 4x wt

Gene/insert name:
4 bulged binding sites for CXCR4 siRNA antisense
Fusion protein or tag:
Rr-luc
Terminal:
N terminal on backbone
Vector backbone:
pRL-TK
(Search Vector Database)
Backbone manufacturer:
Promega
Vector type:
Luciferase
Backbone size w/o insert (bp):
4045
Cloning site 5':
XbaI
Site destroyed during cloning:
No
Cloning site 3':
ApaI
Site destroyed during cloning:
No
5' sequencing primer:
See map List of Sequencing Primers
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Sequence: View sequences (1)
Principal Investigator:
Phil Sharp
Terms and Licenses:
MTA

Comments:

Two original binding sites, separated by 4 nt, were flanked by two of the orginal bulged binding sites, each 11 nt away.

Flanking 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 inner 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.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Specificity of microRNA target selection in translational repression. Doench et al (Genes Dev. 2004 Mar 1. 18(5):504-11. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 11313" in your Materials and Methods section.

Price: $65.00

Available to academic and non-profits only

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This is commonly requested with

11324: pRL-TK let7 A

11325: pRL-TK let7 B

11320: pRL-TK 4x mut G

11312: pGL3 CXCR4 3P

11327: pRL-TK let7 D