pET28 E2 LBD-Tev-E1a-His
(Plasmid #232117)

• Purpose: For bacterial expression of a fusion protein containing from N to C-terminus: the lipoyl-binding domain of Human E2, a TEV protease site, a peptide from Human E1a, and a His tag.
• Depositing Lab: Ruth Sommese
• Publication: Liu et al J Biol Chem. 2023 Mar;299(3):102959. doi: 10.1016/j.jbc.2023.102959. Epub 2023 Jan 28.

Backbone

• Vector backbone: pet28
• Backbone size w/o insert (bp): 5239
• Total vector size (bp): 5635
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: DBT
• Alt name: BCKDHE2
• Alt name: Lipoamide acyltransferase component of branched-chain alpha-keto acid dehydrogenase complex
• Species: H. sapiens (human)
• Insert Size (bp): 403
• Entrez Gene: DBT (a.k.a. BCATE2, BCKAD-E2, BCKADE2, BCKDH-E2, BCOADC-E2, E2, E2B)
• Promoter: T7 Promoter
• Tags / Fusion Proteins:
  • RIGHHSTSDDSSAY (AA331 to 345 (preprocessing, NP_000700.1) of E1a BCKD complex subunit) (C terminal on insert)
  • 6xHis (C terminal on insert)
  • Tev Protease Site (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: T7 Forward
• 3′ sequencing primer: T7 Reverse

Terms and Licenses

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