pSynSRE-Mut-T-Luc
(Plasmid #60490)

• Purpose: Contains a mutant version of the WT pSynSRE-T-Luc reporter plasmid. Four point mutations in the promoter SRE elements abolish SREBP-induced luciferase expression as described by Smith et al., 1988
• Depositing Lab: Timothy Osborne
• Publication: Smith et al J Biol Chem. 1988 Dec 5;263(34):18480-7.

Backbone

• Vector backbone: pGL2 basic
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 5357
• Total vector size (bp): 5544
• Modifications to backbone: Contains mutant version of hamster HMG-CoA synthase promoter sequence harboring SREBP responsive region (-324/-225) fused directly to minimal HMG-CoA sythnase TATA box (-28/+39) that is linked to luciferase coding sequence. This has 4 base pair changes in promoter that make it unresponsive to SREBP
• Vector type: Mammalian Expression, Luciferase

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: HMG-CoA synthase promoter (-324/-225) fused to HMG-CoA synthase TATAA (-28/+39) transcription initiator sequence
• Alt name: 3-Hydroxy-3-methylglutaryl-coenzyme A synthase promoter (partial)
• Species: Hamster
• Insert Size (bp): 187
• Mutation: Four point mutations (G->C or A->C) in the SRE binding regions described by Smith et al., 1988, JBC
• Promoter: Partial HMG-CoA synthase promoter (-324/-225) fused to HMG-CoA Synthase TATAA (-28/+39) transcription initiator
• Tag / Fusion Protein:
  • Luciferase reporter (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SstI or SacI (not destroyed)
• 3′ cloning site: NheI for promoter or HindIII for promoter & TATAA region (not destroyed)
• 5′ sequencing primer: GLprimer1
• 3′ sequencing primer: GLprimer2

Resource Information

• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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