pT2ADW-lox-tdKeima-NES-lox-6011nls(ERK)-P2Av3-RGECO1
(Plasmid
#162486)
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PurposeFor generating transgenic mice expressing ERK biosensor and RGECO1
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Depositing Lab
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Depositing OrganizationKyoto University, Graduate School of Biostudies
Located in Japan -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 162486 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 162486-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 162486-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepT2ADW
- Backbone size w/o insert (bp) 7300
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Modifications to backbonepT2ADW was constructed as follows: The D4Z4 insulator was inserted before the CAG promoter of pT2AL200R175 carrying Tol2 recombination sites. Then, the WPRE sequence of pCSII-EF was inserted before the poly A sequence.
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Vector typeMammalian Expression
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 numberUnknown
Gene/Insert
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Gene/Insert nameERK biosensor based on FRET, and RGECO1
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SpeciesSynthetic
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Insert Size (bp)5000
- Promoter CAG
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site NruI (not destroyed)
- 5′ sequencing primer pCAGGS-5
- (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made bycDNA coding RGECO1 was a gift from Dr. Robert Campbell (Addgene plasmid # 32444 ; http://n2t.net/addgene:32444 ; RRID:Addgene_32444)
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 # 162486-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
-
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.
DNA (Catalog # 162486-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:
pT2ADW-lox-tdKeima-NES-lox-6011nls(ERK)-P2Av3-RGECO1 was a gift from Michiyuki Matsuda (Addgene plasmid # 162486 ; http://n2t.net/addgene:162486 ; RRID:Addgene_162486) -
For your References section:
Visualization of Spatially Controlled Vasospasm by Sympathetic Nerve-Mediated ROCK Activation. Imanishi A, Ichise H, Fan C, Nakagawa Y, Kuwahara K, Sumiyama K, Matsuda M, Terai K. Am J Pathol. 2020 Oct 15. pii: S0002-9440(20)30459-4. doi: 10.1016/j.ajpath.2020.09.012. 10.1016/j.ajpath.2020.09.012 PubMed 33069718
