EcpH-TM
(Plasmid
#85389)
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PurposeExpresses ecliptic pHluorin–ICAM-1 (intercellular adhesion molecule 1) in mammalian cells. Incorporates into viral membrane to label pseudoviruses
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Depositing Lab
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Depositing OrganizationEmory University
Located in United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 85389 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 85389-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 85389-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonep3xFLAG CMV9
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Backbone manufacturerSigma
- Backbone size w/o insert (bp) 6373
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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)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 nameecliptic pHluorin (EcpH) fused to N-terminus of human ICAM-1 transmembrane domain
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SpeciesSynthetic
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Insert Size (bp)900
- Promoter CMV
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Tag
/ Fusion Protein
- 3x FLAG (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (not destroyed)
- 3′ cloning site BamHI (not destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer hGH-pA-R
- (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
A PCR product containing the transmembrane domain of human ICAM-1 was amplified from pCDM8 ICAM-1 (Addgene plasmid # 8632). ecliptic pHluorin (EcpH) was amplified from a Gag-EcpH-expression vector (Sherer et al., Traffic 2003). The EcpH fragment was cloned into p3xFLAG CMV9 using HindIII and EcoRI sites and the ICAM-1 TM fragment was cloned using EcoRI and BamHI.
DNA (Catalog # 85389-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 # 85389-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:
EcpH-TM was a gift from Gregory Melikyan (Addgene plasmid # 85389 ; http://n2t.net/addgene:85389 ; RRID:Addgene_85389) -
For your References section:
Visualization of retrovirus uptake and delivery into acidic endosomes. Miyauchi K, Marin M, Melikyan GB. Biochem J. 2011 Mar 15;434(3):559-69. doi: 10.1042/BJ20101588. 10.1042/BJ20101588 PubMed 21175427
