HA-hDAGLa-V5
(Plasmid
#87674)
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PurposeExpresses human DAGLa protein with a HA tag inserted in the first extracellular loop, and intracellular V5 tag located on the C-terminal
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Depositing Lab
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Depositing OrganizationKing's College London
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 | 87674 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 87674-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 87674-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepcDNA6.2-DEST/V5
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 7341
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Vector typeMammalian Expression
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Selectable markersBlasticidin
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 nameHuman Diacylglycerol Lipase Alpha
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Alt namehDAGLa
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SpeciesH. sapiens (human)
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Insert Size (bp)3177
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MutationHemagglutinin (HA) peptide sequence (YPYDVPDYA) flanked by a glycine-serine-glycine-serine linker on either side was inserted into the first extracellular loop of Human DAGL? sequence
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Entrez GeneDAGLA (a.k.a. C11orf11, DAGL(ALPHA), DAGLALPHA, NSDDR)
- Promoter T7
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Tags
/ Fusion Proteins
- V5 tag (C terminal on backbone)
- HA (N terminal on insert)
Cloning Information
- Cloning method Gateway Cloning
- 5′ sequencing primer T7
- 3′ sequencing primer V5 Rev Priming Site
- (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
Derived from hDAGLa-V5 construct
DNA (Catalog # 87674-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 # 87674-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:
HA-hDAGLa-V5 was a gift from Pat Doherty (Addgene plasmid # 87674 ; http://n2t.net/addgene:87674 ; RRID:Addgene_87674) -
For your References section:
Regulated endosomal trafficking of Diacylglycerol lipase alpha (DAGLalpha) generates distinct cellular pools; implications for endocannabinoid signaling. Zhou Y, Howell FV, Glebov OO, Albrecht D, Williams G, Doherty P. Mol Cell Neurosci. 2016 Oct;76:76-86. doi: 10.1016/j.mcn.2016.08.011. Epub 2016 Sep 3. 10.1016/j.mcn.2016.08.011 PubMed 27595600