pIVM_02
(Plasmid #204500)

• Purpose: Expression of VHL (54-213) for production of VCB complexes in conjunction with plasmid pIVM_26 (EloB 1-104, EloC 17-112)
• Depositing Lab: Alessio Ciulli
• Publication: Cardote et al Structure. 2017 Jun 6;25(6):901-911.e3. doi: 10.1016/j.str.2017.04.009.

Backbone

• Vector backbone: pHAT4
• Backbone manufacturer: Marko Hyvönen, Johan Peränen available from Addgene #112585
• Backbone size w/o insert (bp): 4517
• Total vector size (bp): 5004
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: BL21 or Rosetta at 23°C for protein expression
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: amino acids 54-213 VHL
• Species: H. sapiens (human)
• Insert Size (bp): 485
• Mutation: 0
• GenBank ID: NM 000551.4
• Entrez Gene: VHL (a.k.a. HRCA1, RCA1, VHL1, pVHL)
• Tags / Fusion Proteins:
  • 6xHis (N terminal on insert)
  • Thrombin (N terminal on insert)
  • TEV (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Nco1 (not destroyed)
• 3′ cloning site: EcoR1 (not destroyed)
• 5′ sequencing primer: T7

Terms and Licenses

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

Depositor Comments

Gadd MS, Testa A, Lucas X, Chan KH, Chen W, Lamont DJ, Zengerle M, Ciulli A. Structural basis of PROTAC cooperative recognition for selective protein degradation. Nat Chem Biol. 2017 May;13(5):514-521. doi: 10.1038/nchembio.2329. Epub 2017 Mar 13. PMID: 28288108; PMCID: PMC5392356.