pJH104
(Plasmid #48262)

• Purpose: Template plasmid for PCR-based transplant of Saccharomyces cerevisiae TEF1 promoter in yeast.
• Depositing Lab: Ronald Davis
• Publication: Horecka et al Yeast. 2014 May;31(5):185-93. doi: 10.1002/yea.3006. Epub 2014 Mar 20.

Backbone

• Vector backbone: pBluescript II SK (-)
• Backbone manufacturer: Agilent Technologies
• Backbone size w/o insert (bp): 2961
• Total vector size (bp): 4747
• Vector type: Routine cloning vector

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: TEF1p 5' fragment
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 332
• Mutation: From SK1 background. -460 to -128 relative to TEF1 ORF
• Promoter: TEF1p

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: M13For

Gene/Insert 2

• Gene/Insert name: URA3 5' fragment
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 1126
• Mutation: PCR amplified from pRS316

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: XbaI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: M13For

Gene/Insert 3

• Gene/Insert name: TEF1p 3' fragment
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 343
• Mutation: From SK1 background. -343 to -1 relative to TEF1 ORF
• Promoter: TEF1p

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EagI (not destroyed)
• 5′ sequencing primer: M13Rev

Terms and Licenses

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

Depositor Comments

Created by 4-part ligation with XhoI/EagI cut pBluescript II SK (-). Inserts were XhoI/XbaI TEF1p, XbaI/BamHI URA3, and BamHI/EagI TEF1p.