pcDNA4TO-SCN8A-Variant-1-IRES-mScarlet
(Plasmid #162280)

• Purpose: Eukaryotic expression of human SCN8A variant 1 isoform. This channel has been modified to be stably maintained in standard bacterial strains.
• Depositing Lab: Al George
• Publication: DeKeyser et al J Biol Chem. 2021 Jan-Jun;296:100298. doi: 10.1016/j.jbc.2021.100298. Epub 2021 Jan 15.

Backbone

• Vector backbone: pcDNA4TO-IRES-mScarlet
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 6466
• Total vector size (bp): 12630
• Vector type: Mammalian Expression
• Selectable markers: Zeocin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: SCN8A Variant 1, stabilized
• Alt name: Nav1.6
• Species: H. sapiens (human)
• Insert Size (bp): 6165
• Mutation: Modified human HSPA5 intron 4 inserted after bp2370. IVS intron inserted after bp4370. Silent codon changes at bp4208A>G and bp4211T>C
• GenBank ID: NM_014191
• Entrez Gene: SCN8A (a.k.a. BFIS5, CERIII, CIAT, DEE13, EIEE13, MED, MYOCL2, NaCh6, Nav1.6, PN4)
• Promoter: CMV
• Tag / Fusion Protein:
  • IRES-mScarlet (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: V1-5
• 3′ sequencing primer: pcDNA-R

Resource Information

• Supplemental Documents:
  • pcDNA4TO-SCN8A-Variant-1-IRES-mScarlet.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The SCN8A channel cDNA has been altered to be stably maintained in standard E.coli strains at high copy under standard growth conditions. Please visit https://www.biorxiv.org/content/10.1101/2020.11.05.370429v1 for bioRxiv preprint.