pEGFP-CLCa
(Plasmid
#196912)
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PurposeVisualization of clathrin-coated pit formation in living mammalian cells
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Depositing Lab
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Depositing OrganizationOsaka Metropolitan University
Located in Japan -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 196912 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 196912-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 196912-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepEGFP-C1
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Backbone manufacturerClontech
- Backbone size w/o insert (bp) 4700
- Total vector size (bp) 5600
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Modifications to backboneEcoR I and Sal I sites of pEGFP-ΔC2, which had been derived from pEGFP-C1 by cutting and filling-in Hind III site and converted to C2 reading frame
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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 numberUnknown
Gene/Insert
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Gene/Insert nameClathrin-Coated light chain
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Alt nameNM_007096
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SpeciesH. sapiens (human)
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Insert Size (bp)900
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GenBank ID
- Promoter CMV
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Tag
/ Fusion Protein
- EGFP (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (unknown if destroyed)
- 3′ cloning site SalI (destroyed during cloning)
- 5′ sequencing primer EGFP-C-ter
- 3′ sequencing primer SV40 Poly A site
- (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byObtained by screening the human cDNA library (purchased from Origene).
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
900bp of EcoRI-XhoI fragment of pJG4-5 clone y22-1, from OriGene human MCF7 cDNA Library, was recloned into EcoR I and Sal I sites of pEGFP-ΔC2, which had been derived from pEGFP-C1 by cutting and filling-in Hind III site and converted to C2 reading frame.
DNA (Catalog # 196912-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
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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 # 196912-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:
pEGFP-CLCa was a gift from Kenji Sugimoto (Addgene plasmid # 196912 ; http://n2t.net/addgene:196912 ; RRID:Addgene_196912) -
For your References section:
Rhythmic cycle of clathrin-coated pit formation at the trans-golgi network in human MDA-MB-435 cells. Sakaushi S, Senda-Murata K, Fukada T, Oka S, Sugimoto K. Biosci Biotechnol Biochem. 2007 Feb;71(2):571-4. doi: 10.1271/bbb.60434. Epub 2007 Feb 7. 10.1271/bbb.60434 PubMed 17284835