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Depositing Lab
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Depositing OrganizationJoslin Diabetes Center
Located in the United States of America -
Citations
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Sequence Information
Full plasmid sequence is not available for this item.
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 11238 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 11238-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 11238-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepCMV histidinol
- Backbone size w/o insert (bp) 7583
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Vector typeMammalian Expression
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Selectable markershistidinol
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)XL1 Blue
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameIRS-1
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Alt nameInsulin Receptor Substrate 1
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Alt nameIRS1
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SpeciesH. sapiens (human)
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Insert Size (bp)5800
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MutationC-terminal prenylation motif makes IRS-1 localize to the cell membrane.
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GenBank IDNM_005544
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Entrez GeneIRS1 (a.k.a. HIRS-1)
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Tag
/ Fusion Protein
- CAAX (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site SacI (not destroyed)
- 3′ cloning site SalI (not destroyed)
- 5′ sequencing primer CMV-F
- (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.
Depositor Comments
pCMV human IRS-1 was cut with ApaLI and SalI. A 5' IRS-1 oligo at bp4746 (amino acid 1242) and 3'IRS-1 oligo with 3' IRS-1 sequence fused to the p21ras CAAX prenylation motif (CMSCKCVLS) were used for PCR and the insert was ligated into pCMV human IRS-1. Select cells expressing plasmid with approximately 5mM histidinol (Sigma), depending on the cell type.
DNA (Catalog # 11238-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 # 11238-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:
pCMV human IRS-1 CAAX was a gift from Ronald Kahn (Addgene plasmid # 11238 ; http://n2t.net/addgene:11238 ; RRID:Addgene_11238) -
For your References section:
Cellular compartmentalization in insulin action: altered signaling by a lipid-modified IRS-1. Kriauciunas KM, Myers MG, Kahn CR. Mol Cell Biol. 2000 Sep . 20(18):6849-59. 10.1128/MCB.20.18.6849-6859.2000 PubMed 10958681
Map uploaded by the depositor.