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PurposeThis plasmid contains human codon optimized sequence of mitoLbNOX which is a water-forming NADH oxidase that can be used to increase NAD+/NADH ratio in mammalian cells.
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Depositing Lab
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Depositing OrganizationMassachusetts General Hospital
Located in United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 74448 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 74448-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 74448-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonePUC57
- Backbone size w/o insert (bp) 2710
- Total vector size (bp) 2850
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 namemitoLbNOX
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Alt nameH2O-forming NADH oxidase from Lactobacillus brevis
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SpeciesLactobacillus brevis
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Insert Size (bp)1490
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MutationHuman codon optimized
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Tags
/ Fusion Proteins
- Flag tag with linker (C terminal on insert)
- mitochondrial targeting sequence (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site Not1 (not destroyed)
- 3′ cloning site Mlu1 (not destroyed)
- 5′ sequencing primer GTAAAACGACGGCCAGTG
- 3′ sequencing primer GGAAACAGCTATGACCATG
- (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
In our original publication we express mitoLbNOX in mammalian cells using a Tet3G dox-inducible system from Clontech. We cannot distribute those exact plasmids due to Clontech license restrictions. We recommend that investigators subclone this mitoLbNOX sequence into their preferred expression system.
DNA (Catalog # 74448-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 # 74448-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:
pUC57-mitoLbNOX was a gift from Vamsi Mootha (Addgene plasmid # 74448 ; http://n2t.net/addgene:74448 ; RRID:Addgene_74448) -
For your References section:
Complementation of mitochondrial electron transport chain by manipulation of the NAD+/NADH ratio. Titov DV, Cracan V, Goodman RP, Peng J, Grabarek Z, Mootha VK. Science. 2016 Apr 8;352(6282):231-5. doi: 10.1126/science.aad4017. Epub 2016 Apr 7. 10.1126/science.aad4017 PubMed 27124460