pTH752-CEN-RLuc/min103maxCFLuc
(Plasmid #38218)

• Depositing Lab: Tobias von der Haar
• Publication: Chu et al EMBO J. 2014 Jan 1;33(1):21-34. doi: 10.1002/embj.201385651. Epub 2013 Dec 19.

Backbone

• Vector backbone: pCENBEVY-U
• Backbone size w/o insert (bp): 6616
• Total vector size (bp): 9184
• 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 1

• Gene/Insert name: Firefly Luciferase
• Alt name: FLuc
• Species: Photinus pyralis
• Insert Size (bp): 1644
• Mutation: The first 103 codons of the original FLuc gene were exchanged for those isoaccepting codons decoded by the least abundant tRNAs in yeast. All remaining codons were exchanged for those isoaccepting codons decoded by the most abundant tRNAs. The last three codons of the full-length Firefly luciferase gene have been deleted to yield a luciferase variant which is fully functional, but no longer localises to peroxisomes.
• GenBank ID: AF043450
• Promoter: TDH3 (=GPD)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: M13f

Gene/Insert 2

• Gene/Insert name: Renilla luciferase
• Alt name: RLuc
• Species: Synthetic; Renilla reniformis
• Insert Size (bp): 942
• GenBank ID: AF043450.1
• Promoter: ADH1

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: XmaI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: M13r

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The RLuc gene originated from a plasmid kindly provided by Dr David Bedwell (University of Alabama, Birmingham). Salas-Marco J, Bedwell DM (2004) Molecular and Cellular Biology 24:7769-7778.

Terms and Licenses

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