pET28b-Sad1
(Plasmid #90297)

• Purpose: protein expression
• Depositing Lab: Christine Guthrie
• Publication: Hadjivassiliou et al RNA. 2014 May;20(5):656-69. doi: 10.1261/rna.042838.113. Epub 2014 Mar 28.

Backbone

• Vector backbone: pET28b
• Backbone manufacturer: EMD Biosciences
• Backbone size w/o insert (bp): 5368
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: SAD1
• Alt name: YFR005C
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 1346
• Promoter: T7
• Tag / Fusion Protein:
  • His tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamH1 (unknown if destroyed)
• 3′ cloning site: XhoI (unknown if destroyed)
• 5′ sequencing primer: T7 Fwd

Terms and Licenses

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

Depositor Comments

Full-length Sad1, amino acids 1–448, was cloned from S. cerevisiae S288C into the pET-28b plasmid at the Bam HI and XhoI sites, which fuses a 6-His tag to the N terminus.