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pGEX6P1-optimizedNedd4.2F823D
(Plasmid #165105)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 165105 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pGEX6P1
  • Backbone manufacturer
    GE Healthcare
  • Backbone size w/o insert (bp) 4900
  • 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
    Nedd4.2d F823D
  • Alt name
    NEDD4L
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    2868
  • Mutation
    codon optimized for E. coli expression; F823D; mutation for destabilization of trimer formation
  • Entrez Gene
    NEDD4L (a.k.a. NEDD4-2, NEDD4.2, PVNH7, RSP5, hNEDD4-2)
  • Tag / Fusion Protein
    • GST from plasmid. (N terminal on insert)

Cloning Information

  • Cloning method Ligation Independent Cloning

Terms and Licenses

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

Depositor Comments

Best yields for protein purification were found in BL21 (DE) cells using an overnight induction at 16 degrees C.

5' Cloning Site: BamHI (not destroyed), 3' Cloning Site: EcoRI (not destroyed).

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:

    pGEX6P1-optimizedNedd4.2F823D was a gift from Arthur Haas (Addgene plasmid # 165105 ; http://n2t.net/addgene:165105 ; RRID:Addgene_165105)

  • For your References section:

    Oligomerization of the HECT ubiquitin ligase NEDD4-2/NEDD4L is essential for polyubiquitin chain assembly. Todaro DR, Augustus-Wallace AC, Klein JM, Haas AL. J Biol Chem. 2018 Nov 23;293(47):18192-18206. doi: 10.1074/jbc.RA118.003716. Epub 2018 Oct 4. 10.1074/jbc.RA118.003716 PubMed 30287686
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