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pMM290
(Plasmid #46038)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 46038 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pBI
  • Backbone manufacturer
    Clontech
  • Total vector size (bp) 11707
  • Modifications to backbone
    insertion of polylinker containing Bsp1D1 and XhoI sites
  • Vector type
    Mammalian Expression ; TRE, bidirectional promoter
  • Selectable markers
    Puromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    WRN
  • Alt name
    Werner syndrome, RecQ helicase-like
  • Alt name
    RECQ3
  • Alt name
    RECQL2
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    4514
  • Mutation
    S2G introduced during cloning
  • GenBank ID
    NM_000553.4
  • Entrez Gene
    WRN (a.k.a. RECQ3, RECQL2, RECQL3)
  • Promoter CMV
  • Tag / Fusion Protein
    • 5x myc (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BspD1 (not destroyed)
  • 3′ cloning site Xho1 (not destroyed)
  • 5′ sequencing primer Unknown
  • (Common Sequencing Primers)

Resource Information

  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry

Depositor Comments

Bidirectional vector expresses WRN and GFP at similar expression levels. In the associated article, cells were transiently transfected with this plasmid together with a tTA transactivator plasmid (Gossen, 1992).

This is a mammalian expression vector for catalytically active human WRN protein. lt was constructed from pBI, a Clontech plasmid with a bi-directional promoter+ TRE backbone, modified by insertion of a polylinker containing BspD1 and Xhol sites followed by insertion of myc-WRN WT. Vector backbone is pMM286, myc-WRN WT came from DspD1-Xhol fragment from pMM229.

The WRN sequence in this plasmid contains a number of variations from the WTWRN sequence (NM_000553.4). These polymorphisms are either silent or do not to alter the function of the WRN protein. *The A>G nucleotide change at position 235 was introduced into the WRN sequence during the cloning to generate myc epitope-tagged WRN. The subsequent amino acid change (S2G) does not appear to have any functional consequence on the WRN problem.

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:

    pMM290 was a gift from Raymond Monnat (Addgene plasmid # 46038 ; http://n2t.net/addgene:46038 ; RRID:Addgene_46038)
  • For your References section:

    The Werner syndrome protein has separable recombination and survival functions. Swanson C, Saintigny Y, Emond MJ, Monnat RJ Jr. DNA Repair (Amst). 2004 May 4;3(5):475-82. 10.1016/j.dnarep.2004.01.002 PubMed 15084309