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pETDUET-GST AO7RE Y165A
(Plasmid #139428)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 139428 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pETDUET-1
  • Backbone manufacturer
    Millipore-Sigma
  • Backbone size w/o insert (bp) 5420
  • Total vector size (bp) 5820
  • Modifications to backbone
    An N-terminal GST fusion protein was cloned in place of the His6 tag (NcoI-BamHI). A PreScission Protease (GE Healthcare) sequence (LEVLFQGP) was included to enable the removal of the GST.
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    AO7 aa 126-258
  • Alt name
    RNF25
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    400
  • Mutation
    Changed tyrosine 165 to an alanine
  • GenBank ID
    NM_022453.3
  • Entrez Gene
    RNF25 (a.k.a. AO7)
  • Promoter T7 promoter-1
  • Tag / Fusion Protein
    • GST (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site HindIII (not destroyed)
  • 5′ sequencing primer pET Upstream Primer
  • 3′ sequencing primer DuetDOWN1 Primer
    • (Common Sequencing Primers)

Terms and Licenses

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

    pETDUET-GST AO7RE Y165A was a gift from Allan Weissman (Addgene plasmid # 139428 ; http://n2t.net/addgene:139428 ; RRID:Addgene_139428)

  • For your References section:

    Insights into Ubiquitination from the Unique Clamp-like Binding of the RING E3 AO7 to the E2 UbcH5B. Li S, Liang YH, Mariano J, Metzger MB, Stringer DK, Hristova VA, Li J, Randazzo PA, Tsai YC, Ji X, Weissman AM. J Biol Chem. 2015 Dec 18;290(51):30225-39. doi: 10.1074/jbc.M115.685867. Epub 2015 Oct 16. 10.1074/jbc.M115.685867 PubMed 26475854
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