pMH-macroHC
(Plasmid #82337)

• Purpose: Provides a C-terminal macro domain and 6xHis tag for expression in E. coli
• Depositing Lab: Michael Hothorn
• Publication: Wild et al Protein Sci. 2016 Oct 24. doi: 10.1002/pro.3073.

Backbone

• Vector backbone: pET-24d modified
• Backbone manufacturer: EMBL pepcore
• Backbone size w/o insert (bp): 5502
• Total vector size (bp): 6331
• Modifications to backbone: Insertion of a macro domain fusion tag
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: SPX domain of the Vacuolar Transporter Chaperone 4
• Alt name: VTC4
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 829
• Mutation: Truncated protein, amino acids 1-178
• GenBank ID: NM_001181446
• Entrez Gene: VTC4 (a.k.a. YJL012C, PHM3, YJL012C-A)
• Tags / Fusion Proteins:
  • macro domain of human histone macroH2A1.1 (C terminal on backbone)
  • 6xHis non-cleavable (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XbaI (not destroyed)
• 3′ cloning site: NcoI (not destroyed)
• 5′ sequencing primer: TGCGTCCGGCGTAGAGGATCGAGATCT
• 3′ sequencing primer: CAGCTTCCTTTCGGGCTTTGTTAGCAG

Resource Information

• Supplemental Documents:
  • GenBank file

Terms and Licenses

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