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PurposeKnock eGFP into human VE-cad locus
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Depositing Lab
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Depositing OrganizationUniversity of Wisconsin, Madison
Located in the United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 92309 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 92309-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 92309-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backboneOct4-2A-eGFP backbone
- Backbone size w/o insert (bp) 8563
- Total vector size (bp) 10438
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Modifications to backboneLeft and right arms of Oct4 were replaced with VEcad arms
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Vector typeMammalian Expression, CRISPR
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Selectable markersPuromycin
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 numberUnknown
Gene/Insert
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Gene/Insert nameVEcad Homologous Recombination Arms
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Alt nameCDH5
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SpeciesH. sapiens (human)
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Insert Size (bp)2000
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Mutationabout 30 a.a. truncated at end of protein
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GenBank ID1003
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Entrez GeneCDH5 (a.k.a. 7B4, CD144)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site SbfI, AscI (not destroyed)
- 3′ cloning site NheI, FseI (not destroyed)
- 5′ sequencing primer M13 RVS
- 3′ sequencing primer M13 FWD
- (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
This plasmid functions as described in the related publication, but the 5' homology arm is 528bp instead of ~2kbp.
DNA (Catalog # 92309-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 # 92309-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:
VE-cad-eGFP Donor plasmid was a gift from Sean Palecek (Addgene plasmid # 92309 ; http://n2t.net/addgene:92309 ; RRID:Addgene_92309) -
For your References section:
Human pluripotent stem cell-derived epicardial progenitors can differentiate to endocardial-like endothelial cells. Bao X, Bhute VJ, Han T, Qian T, Lian X, Palecek SP. Bioeng Transl Med. 2017 Jun;2(2):191-201. doi: 10.1002/btm2.10062. Epub 2017 May 22. 10.1002/btm2.10062 PubMed 29170757
Map uploaded by the depositor.
