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pCD-Sub69-NleG6-3 EHEC
(Plasmid #83070)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 83070 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pCDFDuet-1
  • Backbone size w/o insert (bp) 4929
  • Total vector size (bp) 5388
  • Modifications to backbone
    The pCD-Sub vector was constructed based on a pCDF-duet vector. The T7 promoter was substituted with pLtetO-1. The cDHFR fragment (residues 109-187) was PCR-amplified and sub-cloned under the pLtetO1 promoter at the MluI and AscI endonuclease recognition sites (the AscI site was inserted by PCR). A second linker (linker2; as indicated in Figure 4B); was fused to the N-terminus of MBP (Maltose Binding Protein) and cloned into the AscI site. Substrates were PCR-amplified and sub-cloned into SacII and SpeI sites downstream and in-frame with the cDHFR-linker2-MBP. In some vectors the MBP was removed. Some vectors were prepared by complete chemical synthesis assembly (Gibson et al, 2009).
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Spectinomycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    NleG6-3
  • Species
    EHEC
  • Mutation
    WT
  • Tag / Fusion Protein
    • cDHFR- link1-His6-MBP

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:

    pCD-Sub69-NleG6-3 EHEC was a gift from Gali Prag (Addgene plasmid # 83070)

  • 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
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