pWZL Neo Myr Flag PRKAA1
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||20595||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerWilliam Hahn Lab (available at Addgene, #15300)
Vector typeMammalian Expression, Retroviral
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Growth instructionsStbl3 cells to prevent recombination.
Alt nameprotein kinase, AMP-activated, alpha 1 catalytic subunit
SpeciesH. sapiens (human)
Entrez GenePRKAA1 (a.k.a. AMPK, AMPKa1)
/ Fusion Proteins
- Myr (N terminal on backbone)
- Flag (N terminal on backbone)
- Cloning method Gateway Cloning
- 5′ sequencing primer pWZL-Fwd (GAA CCT CCT CGT TCG ACC)
- 3′ sequencing primer pWZL-Rev (TTC CGG GCC CTC ACA TTG) (Common Sequencing Primers)
ORFs were cloned into Gateway compatible pEntry vectors. An LR recombination reaction was performed to move the ORF to pWZL-Neo-Myr-Flag DEST such that the ORF would have 5' myristoylation and flag tags.
This plasmid is part of a kinase library, and the gene has been verified, but has not been fully sequenced for minor mutations. Please see the following link for plasmids from this article that are not part of the kinase library http://www.addgene.org/pubmed/17574021
Addgene's sequencing results suggest that the best reference sequence for the PRKAA1 insert in this plasmid is AB022017.1.
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:pWZL Neo Myr Flag PRKAA1 was a gift from William Hahn & Jean Zhao (Addgene plasmid # 20595 ; http://n2t.net/addgene:20595 ; RRID:Addgene_20595)
For your References section:Integrative Genomic Approaches Identify IKBKE as a Breast Cancer Oncogene (Kinase Library Clones). Boehm JS, Zhao JJ, Yao J, Kim SY, Firestein R, Dunn IF, Sjostrom SK, Garraway LA, Weremowicz S, Richardson AL, Greulich H, Stewart CJ, Mulvey LA, Shen RR, Ambrogio L, Hirozane-Kishikawa T, Hill DE, Vidal M, Meyerson M, Grenier JK, Hinkle G, Root DE, Roberts TM, Lander ES, Polyak K, Hahn WC. Cell. 2007 Jun 15. 129(6):1065-1079. 10.1016/j.cell.2007.03.052 PubMed 17574021