pTH743-CEN-RLuc/slowstaCFLuc
(Plasmid #38223)

• 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): 7122
• Total vector size (bp): 9692
• Modifications to backbone: A mutated GCN4 leader sequence (containing uORF 1 as the sole remaining uORF) was introduced into the BamHI site of this vector. mRNAs expressed from the TDH3 (GPD) promoter therefore contain inefficient 5'-UTRs.
• 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 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 and FLuc genes 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