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PurposeLysosomal pH biosensor consisting of sfGFP-rat Lamp1-mCherry.
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Depositing Lab
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Depositing OrganizationUniversity of California, San Francisco (UCSF)
Located in United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 164477 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 164477-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 164477-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepmCherry-N1
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Backbone manufacturerClontech
- Backbone size w/o insert (bp) 4000
- Total vector size (bp) 6700
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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 nameLAMP1
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SpeciesR. norvegicus (rat)
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Insert Size (bp)2755
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Entrez GeneLamp1 (a.k.a. LGP120)
- Promoter CMV
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Tags
/ Fusion Proteins
- mCherry (C terminal on insert)
- superfolder GFP (N terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer CMV-F, GFP-F, mCherry-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.
DNA (Catalog # 164477-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 # 164477-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:
psfGFP-LAMP1-mCherry (pHLARE) was a gift from Diane Barber (Addgene plasmid # 164477 ; http://n2t.net/addgene:164477 ; RRID:Addgene_164477) -
For your References section:
pHLARE: A New Biosensor Reveals Decreased Lysosome pH in Cancer Cells. Webb BA, Aloisio FM, Charafeddine RA, Cook J, Wittmann T, Barber DL. Mol Biol Cell. 2020 Nov 25:mbcE20060383. doi: 10.1091/mbc.E20-06-0383. 10.1091/mbc.E20-06-0383 PubMed 33237838
