pIRES RED/SP M87C448Y
(Plasmid #89324)

• Purpose: Expression of RFP and M87C448Y 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): 1668
• Mutation: 1-460 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 M87 Kozak sequence was replaced with an optimal Kozak sequence. To detect spastin use an anti-spastin antibody.