prNFL-Q333P-EGFP
(Plasmid #132602)

• Purpose: Expresses EGFP-tagged rat NFL containing the Charcot-Marie-Tooth disease type 2E Q333P mutation
• Depositing Lab: Anthony Brown
• Publication: Stone et al Cytoskeleton (Hoboken). 2019 Jul;76(7-8):423-439. doi: 10.1002/cm.21566. Epub 2019 Nov 6.

Backbone

• Vector backbone: pEGFP-N3
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4698
• Modifications to backbone: Obtained by site-directed mutagenesis of the prNFL-EGFP plasmid. There is a silent mutation at nucleotide 1590G>A.
• 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: High Copy

Gene/Insert

• Gene/Insert name: Nefl
• Alt name: neurofilament protein L
• Species: R. norvegicus (rat)
• Insert Size (bp): 1656
• Mutation: 998A>C, 999G>C; Gln333Pro
• Entrez Gene: Nefl (a.k.a. NF-L, NF68, Nfl)
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: CMV Forward
• 3′ sequencing primer: SV40pA-R

Terms and Licenses

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

Depositor Comments

Note that there is a serine inserted at position 251 in the rat NFL sequence which shifts all amino acids thereafter by one when compared with the human NFL sequence.