URL: http://www.addgene.org/105802
Proper Citation: RRID:Addgene_105802
Bacterial Resistance: Ampicillin
Defining Citation: PMID:29590189
Vector Backbone Description: Backbone Marker:Sammarco & Grabczyk (PMID: 16212928); Vector Backbone:BI-16 (constructed based on pcDNA5/FRT/TO from Invitrogen); Vector Types:Mammalian Expression, Other, Flp-In competent; Bacterial Resistance:Ampicillin
Comments: This vector is a member of the pKK-BI16 series which belongs to the pKK vector family. It enables concomitant expression of two genes, which transcription is driven by a tetracycline responsive bidirectional CMV promoter. This vector is a derivate of the BI16 plasmid (PMID: 16212928) and is suitable for stable cell line generation using the Flp-In system. ORF2 is cloned with the universal SLIC protocol (just three universal PCR primers are required to clone a given coding sequence into all vectors from pKK family by a ligation-independent DNA cloning method), ORF1 can be cloned using SLIC with specifically designed primers (applicable enzymes: Bsp120I, ApaI, MluI, NotI, BspTI, Eco47III, MunI) or conventional restriction-ligase based cloning (see Szczesny RJ et al. for detailed description and protocols; bioRxiv, doi: https://doi.org/10.1101/160101). mClover3 is a brighter derivative of mEGFP. mRuby3 is brighter than mCherry and has slightly shifted excitation and emission spectra. The mClover3-mRuby3 pair is superior to EGFP-mCherry in FRET analysis (PMID: 26879144). Useful for experiments requiring co-expression of two proteins (e.g. FRET measurements).
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Source: Addgene