pREF-GFP kan
(Plasmid #122061)

• Purpose: reporter plasmid for termination and terminator override to express GFP. Marked with URA3
• Depositing Lab: Daniel Reines
• Publication: Reines et al Methods. 2019 Jan 4. pii: S1046-2023(18)30292-5. doi: 10.1016/j.ymeth.2018.12.011.

Backbone

• Vector backbone: pYES2
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5900
• Total vector size (bp): 8700
• Modifications to backbone: kanamycin resistance gene put into ampicillin ORF
• Vector type: Yeast Expression
• Selectable markers: Neomycin (select with G418), URA3

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: intergenic terminator from yeast IMD2
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 100
• GenBank ID: NC_001140.6
• Promoter: GAL1 comes in pYES2

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: PstI (not destroyed)
• 5′ sequencing primer: T7 promoter primer
• 3′ sequencing primer: CYC1 terminator

Gene/Insert 2

• Gene/Insert name: green fluorescent protein
• Species: aequorea victoria
• Insert Size (bp): 1000
• GenBank ID: M62653
• Promoter: GAL1

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: PstI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: CYC1 terminator

Terms and Licenses

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