|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||12565||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 7200
Vector typeMammalian Expression, Luciferase ; Reporter for translation repression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameFour CXCR4 imperfect binding sites
SpeciesH. sapiens (human)
Insert Size (bp)100
Entrez GeneCXCR4 (a.k.a. CD184, D2S201E, FB22, HM89, HSY3RR, LAP-3, LAP3, LCR1, LESTR, NPY3R, NPYR, NPYRL, NPYY3R, WHIM, WHIMS)
/ Fusion Protein
- luciferase (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site XbaI (not destroyed)
- 3′ cloning site ApaI (not destroyed)
- 5′ sequencing primer n/a
- 3′ sequencing primer BGH rev primer (Common Sequencing Primers)
Firefly luciferase was cloned into the HindIII/SpeI sites of pcDNA3.1. Four imperfect complementary binding sites to the antisense strand of CXCR4 siRNA were cloned in the XbaI/ApaI sites in the 3'UTR of luciferase.
Author's sequence data was obtained by using a primer reading forward from inside the luciferase gene.
Imperfect binding sequence is - 5'-AAGTTTTCACAAAGCTAACA
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:PCD FL4X was a gift from Carl Novina (Addgene plasmid # 12565 ; http://n2t.net/addgene:12565 ; RRID:Addgene_12565)
For your References section:Recapitulation of short RNA-directed translational gene silencing in vitro. Wang B, Love TM, Call ME, Doench JG, Novina CD. Mol Cell. 2006 May 19. 22(4):553-60. 10.1016/j.molcel.2006.03.034 PubMed 16713585
Map uploaded by the depositor.