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PurposeMammalian expression vector encoding for the ER targeted and FLAGM1 tagged redox probe roGFPiE (roGFP C147, E147b, C204)
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Depositing Lab
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Depositing OrganizationUniversity of Cambridge
Located in United Kingdom of Great Britain and Northern Ireland -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 47954 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 47954-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 47954-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepcDNA3 derivative
- Backbone size w/o insert (bp) 5370
- Total vector size (bp) 6256
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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 numberHigh Copy
Gene/Insert
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Gene/Insert nameERroGFP
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Speciesjellyfish
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Insert Size (bp)886
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Mutationinsertion E147B
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Tag
/ Fusion Protein
- FLAG (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamH1 (unknown if destroyed)
- 3′ cloning site XhoI (unknown if destroyed)
- 5′ sequencing primer CMV forward
- 3′ sequencing primer SP6
- (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byJames Remington U. Oregon USA
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 # 47954-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 # 47954-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:
ERroGFP_iE_pCDNA3 was a gift from David Ron (Addgene plasmid # 47954 ; http://n2t.net/addgene:47954 ; RRID:Addgene_47954) -
For your References section:
Lifetime imaging of a fluorescent protein sensor reveals surprising stability of ER thiol redox. Avezov E, Cross BC, Kaminski Schierle GS, Winters M, Harding HP, Melo EP, Kaminski CF, Ron D. J Cell Biol. 2013 Apr 15;201(2):337-49. doi: 10.1083/jcb.201211155. 10.1083/jcb.201211155 PubMed 23589496
Map uploaded by the depositor.
