pSMH1741
(Plasmid #160930)

• Purpose: Expression of soluble Sc octamer (H3)
• Depositing Lab: Stephen Harrison
• Publication: Bacterial expression plasmids for S. cerevisiae histone octamers (unpublished)

Backbone

• Vector backbone: pET (https://www.addgene.org/29666/)
• Backbone manufacturer: Berkeley Macrolab
• Backbone size w/o insert (bp): 4786

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Growth instructions: Always pick small colonies. For protein expression, slow-growing cultures will work, but be suspicious of cultures that expand quickly.
• Copy number: Low Copy

Gene/Insert 1

• Gene/Insert name: HHF1
• Alt name: Histone H4
• Species: S. cerevisiae (budding yeast)
• Mutation: codon-optimized for E. coli -- see provided sequence
• Tag / Fusion Protein:
  • 6His-TEV (N terminal on backbone)

Gene/Insert 2

• Gene/Insert name: HTA1
• Alt name: Histone H2A
• Species: S. cerevisiae (budding yeast)
• Tag / Fusion Protein:
  • N-terminal 6His-TEV (N terminal on insert)

Gene/Insert 3

• Gene/Insert name: HTB1
• Alt name: Histone H2B
• Species: S. cerevisiae (budding yeast)
• Tag / Fusion Protein:
  • none

Gene/Insert 4

• Gene/Insert name: HHT1
• Alt name: Histone H3
• Species: S. cerevisiae (budding yeast)
• Tag / Fusion Protein:
  • N-6His-TEV (N terminal on backbone)

Terms and Licenses

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

Depositor Comments

This is an improved version of the plasmid reported here:
https://www.sciencedirect.com/science/article/abs/pii/S096921261930440X
https://doi.org/10.1016/j.str.2019.12.002
All histone genes code for S. cerevisiae proteins. Expression and purification procedures are as described.