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pET28-his-3C_Clu3B6p
(Plasmid #100093)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 100093 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pET28-his-3C
  • Backbone size w/o insert (bp) 5369
  • Total vector size (bp) 6149
  • 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
    14-3-3 sigma human
  • Alt name
    14-3-3 sigma fusion
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    780
  • Mutation
    75-77 residues changed to AAA to facilitate crystallization
  • Entrez Gene
    SFN (a.k.a. YWHAS)
  • Promoter T7
  • Tags / Fusion Proteins
    • HSPB6 peptide (C terminal on insert)
    • His6 (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NcoI (not destroyed)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer T7 promoter
  • 3′ sequencing primer T7 terminator
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

The 3C site was introduced immediately prior to 14-3-3 sequence to facilitate purification by subtractive IMAC.

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:

    pET28-his-3C_Clu3B6p was a gift from Nikolai Sluchanko (Addgene plasmid # 100093 ; http://n2t.net/addgene:100093 ; RRID:Addgene_100093)
  • For your References section:

    Chimeric 14-3-3 proteins for unraveling interactions with intrinsically disordered partners. Sluchanko NN, Tugaeva KV, Greive SJ, Antson AA. Sci Rep. 2017 Sep 20;7(1):12014. doi: 10.1038/s41598-017-12214-9. 10.1038/s41598-017-12214-9 [pii] PubMed 28931924