pCMV-Tag/M1 S245X
(Plasmid
#92366)
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Purposeexpression of M1 S245X spastin in mammalian cells
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 92366 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
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
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, S245X
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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
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/M1 S245X was a gift from Peter Baas (Addgene plasmid # 92366 ; http://n2t.net/addgene:92366 ; RRID:Addgene_92366) -
For your References section:
Studies on truncating mutations of SPAST associated with Hereditary Spastic Paraplegia indicate greater accumulation and toxicity of the M1 isoform of spastin. Solowska JM, Rao AN, Baas PW. Mol Biol Cell. 2017 May 11. pii: mbc.E17-01-0047. doi: 10.1091/mbc.E17-01-0047. 10.1091/mbc.E17-01-0047 PubMed 28495799