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pRL-TK CXCR4 1P
(Plasmid #11309)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 11309 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pRL-TK
  • Backbone manufacturer
    Promega
  • Backbone size w/o insert (bp) 4045
  • Vector type
    Luciferase

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    1 perfect bind site for CXCR4 siRNA antisense
  • Insert Size (bp)
    46
  • Tag / Fusion Protein
    • Rr-luc (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site XbaI (not destroyed)
  • 3′ cloning site ApaI (not destroyed)
  • 5′ sequencing primer See map
  • 3′ sequencing primer EBV-rev
  • (Common Sequencing Primers)

Resource Information

  • Terms and Licenses

Depositor Comments

The end of the ORF and the beginning of the 3' UTR is (with the TAA
stop codon, followed by T, then the XbaI site TCTAGA):
AAATGAACAA TAA T TCTAGA GCGGCCGCT.

We digested the plasmids with XbaI, and ligated in the following:
ctaga ttccgagatatcggtaat gggcc.

This produced a modified vector, still containing an XbaI site, but
now also containing an ApaI site (GGGCCC), to allow for directional
cloning of 3' UTR inserts. The final vector contains, starting with
the stop codon TAA: TAA T Tctaga ttccgagatatcggtaat gggccC TAGA.

All UTR inserts were then constructed in the following format:
TCTAGA CTCGAGCCGG[binding site]ATCGC GGGCCC. A single perfect site
sequence is AAGTTTTCACTCCAGCTAACACCGG.

CXCR4 siRNA used is GUUUUCACUCCAGCUAACACATTCAAAAGUGAGGUCGAUUGU.

How to cite this plasmid ( Back to top)

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 CXCR4 1P was a gift from Phil Sharp (Addgene plasmid # 11309 ; http://n2t.net/addgene:11309 ; RRID:Addgene_11309)
  • For your References section:

    siRNAs can function as miRNAs. Doench JG, Petersen CP, Sharp PA. Genes Dev. 2003 Feb 15. 17(4):438-42. 10.1101/gad.1064703 PubMed 12600936