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Addgene

pCMV-Tag/M1 N184X
(Plasmid #92365)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 92365 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pCMV-Tag
  • Backbone manufacturer
    Stratagene
  • Backbone size w/o insert (bp) 4300
  • Vector type
    Mammalian Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    spastin
  • Alt name
    SPAST
  • Alt name
    NM 014946
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    1929
  • Mutation
    1-194 bp of 5'UTR deleted, N184X
  • Entrez Gene
    SPAST (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

  • Academic/Nonprofit Terms
  • 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

How to cite this plasmid ( Back to top)

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.

  • For your Materials & Methods section:

    pCMV-Tag/M1 N184X was a gift from Peter Baas (Addgene plasmid # 92365 ; http://n2t.net/addgene:92365 ; RRID:Addgene_92365)

  • 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
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