WWP-Luc (p21/WAF1 promoter)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||16451||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector backbonepBluescript II KS (+) + luc
- Backbone size w/o insert (bp) 5600
Vector typeMammalian Expression, Luciferase
Growth in Bacteria
Gene/Insert nameWAF1 promoter
SpeciesH. sapiens (human)
Insert Size (bp)2400
Entrez GeneCDKN1A (a.k.a. CAP20, CDKN1, CIP1, MDA-6, P21, SDI1, WAF1, p21CIP1)
/ Fusion Protein
- luciferase (C terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (unknown if destroyed)
- 3′ cloning site HindIII (unknown if destroyed)
- 5′ sequencing primer T7 (Common Sequencing Primers)
The 2.4kb WAF1 promoter region was obtained by PCR amplification using a P1-WAF1 subclone as template and the primers 5’-CCACAAGCTTCTGACTTCGGCAG-3’and 5’CCCAGGAACAAGCTTGGGCAGCAG-3’. This PCR fragment was cloned into the HindIII site of pBluescript KS(+) (Stratagene) to yield plasmid pWWP. WWP-Luc was cloned by inserting the 2.6kb BamHl luciferase cassette from PG13-Luc into the Xhol sites of pWWP.
Please note that there are a few discrepancies between Addgene's quality control sequence and the top GenBank hit for the promoter sequence. The plasmid should function as reported.
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:WWP-Luc (p21/WAF1 promoter) was a gift from Bert Vogelstein (Addgene plasmid # 16451 ; http://n2t.net/addgene:16451 ; RRID:Addgene_16451)
For your References section:WAF1, a potential mediator of p53 tumor suppression. el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B. Cell. 1993 Nov 19. 75(4):817-25. 10.1016/0092-8674(93)90500-P PubMed 8242752