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PurposeContains 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
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Depositing Lab
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Depositing OrganizationUniversity of California, Irvine
Located in United States of America -
Publication
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Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 60490 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 60490-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 60490-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepGL2 basic
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Backbone manufacturerPromega
- Backbone size w/o insert (bp) 5357
- Total vector size (bp) 5544
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Modifications to backboneContains 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
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Vector typeMammalian Expression, Luciferase
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameHMG-CoA synthase promoter (-324/-225) fused to HMG-CoA synthase TATAA (-28/+39) transcription initiator sequence
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Alt name3-Hydroxy-3-methylglutaryl-coenzyme A synthase promoter (partial)
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SpeciesHamster
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Insert Size (bp)187
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MutationFour 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
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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)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
DNA (Catalog # 60490-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 60490-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
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.
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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