prNFL-EGFP
(Plasmid #132605)

• Purpose: Expresses EGFP-tagged rat NFL
• 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: PCR was used to amplify the NFL cDNA without the C-terminal myc tag using the pNFL-myc plasmid as the template. The amplified sequence was then subcloned into the multiple cloning site of the pEGFP-C1 vector, generating the following linker sequence: 5'-GGATCCATCGCCACC-3'. 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
• 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

Resource Information

• Supplemental Documents:
  • prNFL-EGFP.gb
• A portion of this plasmid was derived from a plasmid made by: The prNFL-EGFP fluorescent fusion construct was created from a rat pNFL-myc construct provided by Chris Miller (Yates et al., Eur. J. Cell Biol. 88:193-202, 2009).

Terms and Licenses

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