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

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 60490 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

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
  • 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
  • (Common Sequencing Primers)

Resource Information

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pSynSRE-Mut-T-Luc was a gift from Timothy Osborne (Addgene plasmid # 60490 ; http://n2t.net/addgene:60490 ; RRID:Addgene_60490)
  • For your References section:

    Multiple sterol regulatory elements in promoter for hamster 3-hydroxy-3-methylglutaryl-coenzyme A synthase. Smith JR, Osborne TF, Brown MS, Goldstein JL, Gil G. J Biol Chem. 1988 Dec 5;263(34):18480-7. PubMed 2903862