pORF1-Flag
(Plasmid #190565)

• Purpose: Mammalian expression plasmid encoding the first open reading frame (ORF1, expressing the protein ORF1p) of human L1 (L1.3); carries a C-terminus Flag tag.
• Depositing Lab: Anthony Furano
• Publication: Cook et al Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):4298-303. doi: 10.1073/pnas.1416869112. Epub 2015 Mar 23.

Backbone

• Vector backbone: pcDNA 3.1(+)-puro
• Backbone manufacturer: ThermoFisher
• Backbone size w/o insert (bp): 5159
• Total vector size (bp): 6200
• Modifications to backbone: Neomycin resistance gene was removed and replaced with puromycin resistance gene for selection in mammalian cells.
• Vector type: Mammalian Expression
• Selectable markers: Puromycin

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: ORF1 (from L1.3)
• Alt name: p40
• Species: H. sapiens (human)
• Insert Size (bp): 1041
• Promoter: CMV
• Tag / Fusion Protein:
  • FLAG (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • pORF1-Flag SnapGene annotated sequence
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

The bases GCA were inserted between BamHI and the first codon of ORF1 (ATG) for a consensus Kozak sequence as described in the Supporting Information in Cook, Jones, and Furano, 2015, 10.1073/pnas.1416869112, PMID: 25831499.