pCMV-MCS-eDHFR(69K6)-EGFP
(Plasmid
#172101)
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PurposeMammalian expression of a protein of interest fused to the N-terminus of eDHFR(69K6)-EGFP
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Depositing Lab
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Depositing OrganizationNagoya Institute of Technology
Located in Japan -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 172101 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 172101-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 172101-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepEGFP-N1
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Backbone manufacturerClontech
- Backbone size w/o insert (bp) 4695
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Vector typeMammalian Expression
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Selectable markersNeomycin (select with G418)
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 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 name22aa-eDHFR(69K6)-EGFP
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SpeciesSynthetic
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Insert Size (bp)605
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MutationeDHFR(69K6): Hexalysine (K6) sequence is inserted between Asp69 and Asp70 of E. coli DHFR
- Promoter CMV
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (not destroyed)
- 3′ cloning site BamHI (not destroyed)
- 5′ sequencing primer unknown
- (Common Sequencing Primers)
Resource Information
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
The chemical mDcTMP, to control protein translocation, can be obtained from Funakoshi Co., Ltd. https://www.funakoshi.co.jp/exports_contents/95015. Please visit https://www.biorxiv.org/content/10.1101/2021.03.16.435568v1 for bioRxiv preprint.
DNA (Catalog # 172101-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 # 172101-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-MCS-eDHFR(69K6)-EGFP was a gift from Shinya Tsukiji (Addgene plasmid # 172101 ; http://n2t.net/addgene:172101 ; RRID:Addgene_172101) -
For your References section:
A chemogenetic platform for controlling plasma membrane signaling and synthetic signal oscillation. Suzuki S, Nakamura A, Hatano Y, Yoshikawa M, Yoshii T, Sawada S, Atsuta-Tsunoda K, Aoki K, Tsukiji S. Cell Chem Biol. 2022 Sep 15;29(9):1446-1464.e10. doi: 10.1016/j.chembiol.2022.06.005. Epub 2022 Jul 13. 10.1016/j.chembiol.2022.06.005 PubMed 35835118
