pIRES RED/SP M1C448Y
(Plasmid #89323)

• Purpose: Expression of RFP and M1C448Y spastin in mammalian cells
• Depositing Lab: Peter Baas
• Publication: Solowska et al J Neurosci. 2014 Jan 29;34(5):1856-67. doi: 10.1523/JNEUROSCI.3309-13.2014.

Backbone

• Vector backbone: pIRES2 DsRED-Express2
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 5265
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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
• Entrez Gene: SPAST (a.k.a. ADPSP, FSP2, SPG4)
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Eco47III (destroyed during cloning)
• 3′ cloning site: BglII (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: pCDH-rev

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The endogenous M1 Kozak sequence was replaced with an optimal Kozak sequence to prevent leaky translation of M87 spastin. To detect spastin use an anti-spastin antibody.