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URL: http://www.addgene.org/105779
Proper Citation: RRID:Addgene_105779
Bacterial Resistance: Ampicillin
Defining Citation: PMID:29590189
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pcDNA5/FRT/TO; Vector Types:Mammalian Expression, Flp-In competent; Bacterial Resistance:Ampicillin
Comments: This vector is a member of the pKK vector family. Its cloning site was optimized for the SLIC approach, however, a traditional restriction and ligation based approach is also possible (see Szczesny RJ et al. for detailed protocols; bioRxiv, doi: https://doi.org/10.1101/160101). This vector is a derivate of the pcDNA5/FRT/TO plasmid (Invitrogen) and is suitable for stable cell line generation using the Flp-In system. Transcription of the gene of interest is driven by a tetracycline responsive CMV promoter. A given coding sequence can be cloned into all pKK vectors using just three universal PCR primers and a ligation-independent DNA cloning method. mCherry descends from the first truly monomeric red fluorescent protein mRFP1, which was obtained by mutating DsRed form Discosoma sp. Note that EGFP and mCherry come from different organisms and their sequences differ greatly. However, GFP-type termini were introduced into mCherry to increase tolerance of N- and C-terminal fusions in mammalian cells (PMID: 15558047). Thus, terminal sequences of EGFP and mCherry are the same.
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Source: Addgene
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