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Addgene

pWZL-Neo-Myr-Flag-DEST
(Plasmid #15300)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 15300 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Cloning Grade DNA 15300-DNA.cg 2 µg of cloning grade DNA in Tris buffer 1 $105
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Backbone

  • Vector backbone
    pWZL-Neo-Myr-Flag-DEST
  • Vector type
    Mammalian Expression, Retroviral
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol and Ampicillin, 25 & 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    ccdB Survival
  • Growth instructions
    DB3.1, ccdB Survival
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    None
  • Tags / Fusion Proteins
    • Myr (N terminal on backbone)
    • Flag (N terminal on backbone)

Cloning Information

  • 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)

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

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.

LR reaction:
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.

Information for Cloning Grade DNA (Catalog # 15300-DNA.cg) ( Back to top)

Purpose

Cloning grade DNA is suitable for use in PCR, cloning reactions, or transformation into E. coli. The purity and amount is not suitable for direct transfections.

Delivery

  • Amount 2 µg
  • Guaranteed Concentration 100 ng/µl +/- 5 ng/µl
  • Pricing $105 USD
  • Storage DNA can be stored at 4℃ (short term) or -20℃ (long term).

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Quality Control

Addgene has verified this plasmid using Next Generation Sequencing. Results are available here

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:

    pWZL-Neo-Myr-Flag-DEST was a gift from Jean Zhao (Addgene plasmid # 15300 ; http://n2t.net/addgene:15300 ; RRID:Addgene_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
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