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PurposemCherry tagged Geph for mammalian expression
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Depositing Lab
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Depositing OrganizationUniversity of Zurich
Located in Switzerland -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 68820 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 68820-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 68820-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepEGFP C2
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Backbone manufacturerClontech
- Backbone size w/o insert (bp) 4700
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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 nameGephyrin
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Alt nameGPHN
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SpeciesR. norvegicus (rat)
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Insert Size (bp)2200
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Mutationsilent mutations on internal EcoRI site. Gephyrin D566N (see depositor comment below)
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GenBank IDNM022865.3
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Entrez GeneGphn (a.k.a. Geph)
- Promoter CMV
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Tag
/ Fusion Protein
- mCherry (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site KpnI (not destroyed)
- 5′ sequencing primer EGFP-C
- (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
EGFP was removed using NheI and XhoI. mCherry was amplified and inserted using NheI and XhoI sites. Gephyrin D566N mutation is a polymorphism and is not a concern for plasmid function.
DNA (Catalog # 68820-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 # 68820-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:
pmCherryC2-Gephyrin P1 was a gift from Shiva Tyagarajan (Addgene plasmid # 68820 ; http://n2t.net/addgene:68820 ; RRID:Addgene_68820) -
For your References section:
Regulation of GABAergic synapse formation and plasticity by GSK3beta-dependent phosphorylation of gephyrin. Tyagarajan SK, Ghosh H, Yevenes GE, Nikonenko I, Ebeling C, Schwerdel C, Sidler C, Zeilhofer HU, Gerrits B, Muller D, Fritschy JM. Proc Natl Acad Sci U S A. 2011 Jan 4;108(1):379-84. doi: 10.1073/pnas.1011824108. Epub 2010 Dec 20. 10.1073/pnas.1011824108 PubMed 21173228
