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URL: http://www.addgene.org/105793
Proper Citation: RRID:Addgene_105793
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. Enhanced green florescent protein (EGFP) is a mutant of wild-type GFP with humanized sequence and mutations (F64L, S65T) that improve its spectral properties (PMID: 8805248). EGFP is not truly monomeric in contrast to mEGFP (a property conferred by the A206K mutation, PMID: 11988576, PMID: 22869113).
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Source: Addgene
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