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pGEN25-Ub Hs
(Plasmid #83232)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 83232 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pHIS parallele 2
  • Backbone size w/o insert (bp) 5289
  • Total vector size (bp) 5780
  • Modifications to backbone
    The pND-Ub vector is based on a modified pZE21 vector, contacting the pLtetO-1. cDNAs were amplified by polymerase chain reaction (PCR), digested and sub-cloned into the vectors. The mouse cDNA of the nDHFR fragment (residues 1-108) was PCR-amplified and sub-cloned between the KpnI and PacI sites (the PacI site was inserted into the vector by PCR) into the pZE21 vector. Then a flexible linker (Figure 4) was constructed a using two-step PCR method and digested with PacI and NotI (the NotI site was inserted into the vector by PCR) and cloned into the vector downstream to the pLtetO1-nDHFR. Ubiquitylation apparatus cassettes containing a His6-Ub-E2-E1 were PCR-amplified from pGEN plasmids expressing different E2s or Ub mutants (Keren-Kaplan et al, 2011) and sub-cloned as fusion to the C-terminus of the nDHFR-linker1 into the NotI and AvrII endonuclease recognition sites.
  • Vector type
    Bacterial Expression

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    Ubiqutin
  • Alt name
    Ubq
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    231
  • Entrez Gene
    UBC (a.k.a. HMG20)
  • Tag / Fusion Protein
    • His (N terminal on insert)

Resource Information

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

    pGEN25-Ub Hs was a gift from Gali Prag (Addgene plasmid # 83232 ; http://n2t.net/addgene:83232 ; RRID:Addgene_83232)
  • For your References section:

    A bacterial genetic selection system for ubiquitylation cascade discovery. Levin-Kravets O, Tanner N, Shohat N, Attali I, Keren-Kaplan T, Shusterman A, Artzi S, Varvak A, Reshef Y, Shi X, Zucker O, Baram T, Katina C, Pilzer I, Ben-Aroya S, Prag G. Nat Methods. 2016 Nov;13(11):945-952. doi: 10.1038/nmeth.4003. Epub 2016 Oct 3. 10.1038/nmeth.4003 PubMed 27694912