pcDNA3-P1-LoxP-Neo-N-miRFP670-C
(Plasmid #159434)

• Purpose: CRISPR knock-in donor construct with fluorescent reporter and removable selectable marker
• Depositing Lab: Jianhua Xing
• Publication: Chen et al Biol Methods Protoc. 2020 Mar 20;5(1):bpaa006. doi: 10.1093/biomethods/bpaa006. eCollection 2020.

Backbone

• Vector backbone: pcDNA3
• Total vector size (bp): 7988
• Vector type: CRISPR
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: Neomycin
• Insert Size (bp): 1490
• Promoter: SV40
• Tags / Fusion Proteins:
  • 5' linker pair 1 (N terminal on backbone)
  • LoxP (N terminal on insert)
  • LoxP (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: AgeI (not destroyed)
• 3′ cloning site: n/a (unknown if destroyed)
• 5′ sequencing primer: T7

Gene/Insert 2

• Gene/Insert name: miRFP670
• Insert Size (bp): 945
• Tags / Fusion Proteins:
  • N terminal peptide linker (N terminal on insert)
  • C terminal peptide linker (C terminal on insert)
  • 3' linker pair 1 (C terminal on backbone)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: n/a (unknown if destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: GAAAGGACAGTGGGAGTGGCACCTTCCAGGGCT

Terms and Licenses

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