pNB789
(Plasmid #60166)

• Purpose: Firefly luciferase (Promega) yeast codon-optimized yellow fluorescent protein (yEVenus) fusion with PEST degron driven by RNR1 promoter.
• Depositing Lab: Nicolas Buchler
• Publication: Mazo-Vargas et al Mol Biol Cell. 2014 Sep 17. pii: mbc.E14-07-1187.

Backbone

• Vector backbone: p406
• Backbone size w/o insert (bp): 5820
• Total vector size (bp): 8739
• Vector type: Yeast Expression
• Selectable markers: URA3

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: FLuc-yEVenus-PEST
• Alt name: Firefly luciferase and yeast codon-optimized yellow fluorescent protein fusion with PEST degron.
• Alt name: p406-RNR1pr-FLuc-yEVenus-PEST
• Species: Photinus pyralis
• Insert Size (bp): 2919
• Mutation: FLuc has A4V & S504G (no functional effect)
• Promoter: RNR1pr
• Tag / Fusion Protein:
  • PEST degron (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: PacI (not destroyed)
• 3′ cloning site: AscI (not destroyed)
• 5′ sequencing primer: cactttatgcttccggctcctatgtt
• 3′ sequencing primer: CTGCCGGTAGAGGTGTGGTCAATAAG

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry