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. 2015 Apr 9;10(4):e0123646. doi: 10.1371/journal.pone.0123646

Fig 3. Overview of the system and plasmid combinations.

Fig 3

Two Entry vectors suitable for C-terminal (left) or N-terminal (right) tagging are modified with genes of interest (GOI1 = red, GOI2 = green). The two resulting Entry clones are combined in a MultiSite Gateway LR reaction with an application-specific 3rd Entry clone and Destination vector (below) resulting in the desired Expression clone (bottom). Plasmids pWRG-B2H-ENTR-N and pWRG-B2H-DEST-N allow for N-terminal tagging in bacterial two hybrid (B2H) applications if used together with compatible pWRG-ENTR-N1/2-derived Entry clones (right). Boxes represent the genes or gene fragments as indicated (gluc 105 = GlucM18–105, gluc 106 = Gluc106–185, scfp3a = Super CFP 3a, syfp2 = Super YFP 2, snap = SNAP-tag, halo = HaloTag 7, t18 = T18 fragment of CyaA from B. pertussis, t25 = T25 fragment of CyaA from B. pertussis), black rectangles symbolize stop-codons, tetracycline-inducible promoters are depicted as arrows and Ω stand for rrnB-derived terminators. Resistance cassettes, attR- and attL sites as well as by-products of the reactions are not shown.