pET28-his-3C_Clu3B6p
(Plasmid #100093)

• Purpose: expresses easily crystallizable engineered 14-3-3 sigma variant fused to the HSPB6 phosphorylatable peptide
• Depositing Lab: Nikolai Sluchanko
• Publication: Sluchanko et al Sci Rep. 2017 Sep 20;7(1):12014. doi: 10.1038/s41598-017-12214-9.

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, 50 μg/mL
• 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

Resource Information

• Supplemental Documents:
  • Clu3-B6p_insert.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

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