Pcdna5 frt plasmid
The PcDNA5/FRT plasmid is a vector designed for stable transfection and transgene expression in mammalian cells. It contains an FRT site for Flp-mediated recombination and a hygromycin resistance gene for selection. This plasmid serves as a tool for genetic engineering and recombination studies in cell line development.
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11 protocols using pcdna5 frt plasmid
Doxycycline-Inducible N2A Flp-In Stable Lines
Cloning and Expression of Protein Constructs
Purification and Analysis of IgG1 Antibodies
Molecular Cloning of Protein Constructs
Molecular Engineering of Fluorescent AQP3 and TMC1
To express TMC1-YFP construct, the C-terminus of coding region of mouse Tmc1 was fused to the coding sequence of mVenus reporter using Hind III and BamHI sites in an mVenus-N1 vector36 (link). Sequencing of this construct confirmed the identity as murine Tmc1 (Pumbed NM_028953).
Cloning and Expressing Human α1 and α2
Stable Cell Line Generation of Ndc80 Tail Mutants
Generation of AQP6-tGFP Fusion Constructs
The AQP6-tGFP construct was subcloned into a pcDNA5/FRT plasmid (Invitrogen, Carlsbad, CA) using restriction enzymes BamH1/EcoRV while the AQP6 was subcloned using Hpa1/EcoRI into a pcDNA5/FRT plasmid containing tGFP. Each construct was custom synthesized by Genscript, codon optimized for mammalian expression in HEK293 cells. Each AQP6-based sequence was then subcloned into the AQP3-tGFP-pcDNA5/FRT plasmid using Hpa1/EcoRV restriction sites. All constructs were confirmed by sequencing.
Generation of pDF_FRT Plasmid
The complete vector sequences are available upon request.
Stable Expression of Human Alpha Subunits
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