pT2ADW-lox-mCherryNLS-pA-lox-4493NES
(Plasmid
#138374)
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PurposeBooster-PKA for the generation of mice
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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 | 138374 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 138374-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 138374-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepT2ADW
- Backbone size w/o insert (bp) 6500
- Total vector size (bp) 11242
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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 namelox-mCherry-lox-Booster-PKA
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SpeciesS. cerevisiae (budding yeast), Synthetic
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Insert Size (bp)2600
- Promoter CAG
Cloning Information
- Cloning method Gateway Cloning
- 5′ sequencing primer pCAGGS-5
- (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
Depositor found that the linker region of the Booster-PKA insert may be 42 bp longer than shown in their reference sequence, but that the plasmid functions as reported in the associated publication.
Sequences are based on next-generation sequencing (NGS) results where indicated (Addgene NGS Result), or assembled from reference sequences and/or Sanger results (Addgene Assembled Sequence). Sequence provided by depositing laboratory may be theoretical/predicted or based on Sanger/NGS sequencing results. Discrepancies between sequencing results obtained by Addgene and the original sequence provided by the depositor may be present.
DNA (Catalog # 138374-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 # 138374-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-mCherryNLS-pA-lox-4493NES was a gift from Michiyuki Matsuda (Addgene plasmid # 138374 ; http://n2t.net/addgene:138374 ; RRID:Addgene_138374) -
For your References section:
Booster, a Red-Shifted Genetically Encoded Forster Resonance Energy Transfer (FRET) Biosensor Compatible with Cyan Fluorescent Protein/Yellow Fluorescent Protein-Based FRET Biosensors and Blue Light-Responsive Optogenetic Tools. Watabe T, Terai K, Sumiyama K, Matsuda M. ACS Sens. 2020 Feb 26. doi: 10.1021/acssensors.9b01941. 10.1021/acssensors.9b01941 PubMed 32101394