pCMV-Tag/M1C448Y
(Plasmid
#89319)
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Purposeexpression of M1 C448Y spastin in mammalian cells
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Depositing Lab
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Depositing OrganizationDrexel University
Located in the United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 89319 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 89319-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 89319-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepCMV-Tag
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Backbone manufacturerStratagene
- Backbone size w/o insert (bp) 4300
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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 namespastin
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Alt nameSPAST
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Alt nameNM 014946
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SpeciesH. sapiens (human)
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Insert Size (bp)1929
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Mutation1-194 bp of 5'UTR deleted, C448Y
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Entrez GeneSPAST (a.k.a. ADPSP, FSP2, SPG4)
- Promoter CMV
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoICR I/EcoRV (destroyed during cloning)
- 3′ cloning site BamH I/Bgl II (destroyed during cloning)
- 5′ sequencing primer T3
- 3′ sequencing primer 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
Cloning of spastin cDNA into pCMV-Tag EcoICR I and BamH I sites removed myc tag. To detect spastin use anti-spastin antibody.The endogenous M1 Kozak sequence was replaced with an optimal Kozak sequence to prevent leaky translation of M87 spastin.
DNA (Catalog # 89319-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 # 89319-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:
pCMV-Tag/M1C448Y was a gift from Peter Baas (Addgene plasmid # 89319 ; http://n2t.net/addgene:89319 ; RRID:Addgene_89319) -
For your References section:
Pathogenic mutation of spastin has gain-of-function effects on microtubule dynamics. Solowska JM, D'Rozario M, Jean DC, Davidson MW, Marenda DR, Baas PW. J Neurosci. 2014 Jan 29;34(5):1856-67. doi: 10.1523/JNEUROSCI.3309-13.2014. 10.1523/JNEUROSCI.3309-13.2014 PubMed 24478365
