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pGEX-optimized E6AP F727D
(Plasmid #165101)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 165101 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pGEX4T1
  • Backbone manufacturer
    GE Healthcare
  • Backbone size w/o insert (bp) 4900
  • Vector type
    Bacterial Expression

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    E6AP F727D
  • Alt name
    UBE3A
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    2620
  • Mutation
    codon optimized for E. coli expression
  • Entrez Gene
    UBE3A (a.k.a. ANCR, AS, E6-AP, EPVE6AP, HPVE6A)
  • 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

Depositor Comments

Amino acid numbering is based on Isoform 1 though this clone is isoform 3. The sequence is 5'-SPLKYLFRPEE-3' where F has been mutated to D. Best yields for protein purification were found in BL21 (DE) cells using an overnight induction at 16 degrees C.

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

    pGEX-optimized E6AP F727D was a gift from Arthur Haas (Addgene plasmid # 165101 ; http://n2t.net/addgene:165101 ; RRID:Addgene_165101)
  • For your References section:

    The active form of E6-associated protein (E6AP)/UBE3A ubiquitin ligase is an oligomer. Ronchi VP, Klein JM, Edwards DJ, Haas AL. J Biol Chem. 2014 Jan 10;289(2):1033-48. doi: 10.1074/jbc.M113.517805. Epub 2013 Nov 22. 10.1074/jbc.M113.517805 PubMed 24273172