Purpose(Empty Backbone) a Gateway-compatible retroviral destination vector which adds a myristoylation sequence and a FLAG tag to each introduced ORF
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||15300||Plasmid sent as bacteria in agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector typeMammalian Expression, Retroviral
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Bacterial Resistance(s)Chloramphenicol and Ampicillin
Growth Strain(s)ccdB Survival
Growth instructionsDB3.1, ccdB Survival
/ Fusion Proteins
- Myr (N terminal on backbone)
- Flag (N terminal on backbone)
- Cloning method Gateway Cloning
- 5′ sequencing primer GAA CCT CCT CGT TCG ACC
- 3′ sequencing primer TTC CGG GCC CTC ACA TTG (Common Sequencing Primers)
pWZL-Neo-Myr-Flag-DEST (pWN-MF-DEST) is a Gateway-compatible retroviral destination vector which adds a myristoylation sequence and a FLAG tag to each introduced ORF.
Created by inserting an N-terminal myr-FLAG tag and the Gateway-selectable cassette into the pWzl-Neo retroviral vector. To create specific pWN-MF-ORF vectors, perform site-specific LR recombination reactions.
150ng DEST vector + 50ng ENTRY vector + 10xLR Buffer + LR enzyme + dH20. Incubate at 25oC for 1 hour. Add 1uL Proteinase K (20mg/ml), and incubate at 37oC for 10min. Transform into DH5alpha cells. See Invitrogen's website for detailed protocols and for LR enzyme.
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-DEST was a gift from Jean Zhao (Addgene plasmid # 15300)
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