pcDNA3.1(+)/Luc2=tdT
(Plasmid #32904)

• Purpose: Expresses a fusion of firefly luciferase and tandem Tomato red fluorescent protein in mammalian cells.
• Depositing Lab: Christopher Contag
• Publication: Patel et al Cancer Res. 2010 Dec 15;70(24):10141-9.

Backbone

• Vector backbone: pcDNA3.1(+)
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5428
• Modifications to backbone: The luc2=tdT fusion gene was created by PCR amplification of the Luc2 gene (pGL4.10, Promega, Madison, WI) and cloned by restriction digestion upstream of the tdTomato gene in the vector pRSET-B-tdTomato (kindly provided by Roger Tsien, UC San Diego). The luc2=tdT fusion gene was excised with EcoRI and inserted into the EcoRI site in the pcDNA3.1(+) multiple cloning site.
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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: luc2
• Species: Photonis pyralis (firefly)
• Insert Size (bp): 3129
• GenBank ID: AY738222
• Promoter: CMV-IE
• Tag / Fusion Protein:
  • tdTomato red fluorescent protein (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: n/a

Gene/Insert 2

• Gene/Insert name: tandem Tomato Red Fluorescent Protein
• Species: Discosoma
• GenBank ID: AY678269
• Promoter: n/a

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: pcDNA3.1/BGH reverse primer

Resource Information

• A portion of this plasmid was derived from a plasmid made by: This luc2=tdT (luciferase-tandem Tomato Red Fluorescent Protein) fusion gene was cloned by the submitter, Dr. Michael H. Bachmann, Dept. of Pediatrics, Stanford University, Stanford, CA 94305-5427.
• Articles Citing this Plasmid:
  • 22 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry