B-GRAM-H
(Plasmid
#211706)
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PurposeExpression of a dimerization-dependent fluorescent protein (ddFP) subunit (B) fused with the GRAM domain of GRAMD1b carrying a G187L mutation (B-GRAM-H) in mammalian cells
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Depositing Lab
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Depositing OrganizationNanyang Technological University
Located in Singapore -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 211706 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 211706-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 211706-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepEGFP-C1
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Backbone manufacturerClontech
- Total vector size (bp) 5035
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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 numberHigh Copy
Gene/Insert
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Gene/Insert nameB-tagged GRAM domain of GRAMD1b/Aster-B
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Alt nameB-GRAM1b
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SpeciesH. sapiens (human)
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Insert Size (bp)1072
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MutationChanged Glycine 187 to Leucine
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GenBank IDNM_020716.4
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Entrez GeneGRAMD1B (a.k.a. LINC01059)
- Promoter CMV
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Tag
/ Fusion Protein
- ddFP-B3 (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NheI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer CMV Forward
- 3′ sequencing primer SV40pA-R
- (Common Sequencing Primers)
Resource Information
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 # 211706-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 # 211706-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:
B-GRAM-H was a gift from Yasunori Saheki (Addgene plasmid # 211706 ; http://n2t.net/addgene:211706 ; RRID:Addgene_211706) -
For your References section:
Visualization of accessible cholesterol using a GRAM domain-based biosensor. Koh DHZ, Naito T, Na M, Yeap YJ, Rozario P, Zhong FL, Lim KL, Saheki Y. Nat Commun. 2023 Oct 25;14(1):6773. doi: 10.1038/s41467-023-42498-7. 10.1038/s41467-023-42498-7 PubMed 37880244