hHS1-M7A
(Plasmid
#159170)
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PurposeHuman codon-optimized cytosolic labile heme reporter
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Depositing Lab
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Depositing OrganizationGeorgia Institute of Technology
Located in the United States of America -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 159170 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 159170-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 159170-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepcDNA3-eGFP
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Backbone manufacturerDoug Golenbock (Addgene #13031)
- Backbone size w/o insert (bp) 6146
- Total vector size (bp) 7906
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Vector typeMammalian Expression
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Selectable markersBleoR
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 namehHS1-M7A
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SpeciesH. sapiens (human)
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Insert Size (bp)1760
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MutationMutated Met 7 of the cyt b562 module of hHS1 to Ala (M7A), creates medium affinity heme sensor
- Promoter CMV
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site XhoI (not destroyed)
- 5′ sequencing primer CMV-F or T7
- (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
The first-generation heme sensor, HS1, consists of a heme-binding domain, the His/Met coordinating 4-alpha-helical bundle hemoprotein cytochrome b562 (Cyt b562) (13), fused to a pair of fluorescent proteins, EGFP and Katushka 2 (mKATE2)
DNA (Catalog # 159170-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
-
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 # 159170-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:
hHS1-M7A was a gift from Amit Reddi (Addgene plasmid # 159170 ; http://n2t.net/addgene:159170 ; RRID:Addgene_159170) -
For your References section:
Heme dynamics and trafficking factors revealed by genetically encoded fluorescent heme sensors. Hanna DA, Harvey RM, Martinez-Guzman O, Yuan X, Chandrasekharan B, Raju G, Outten FW, Hamza I, Reddi AR. Proc Natl Acad Sci U S A. 2016 Jul 5;113(27):7539-44. doi: 10.1073/pnas.1523802113. Epub 2016 May 31. 10.1073/pnas.1523802113 PubMed 27247412
