Figure 3.

Choice of module plasmids for expression of FP-GOI. (a) Detailed construct of an integration plasmid for expression of a GFP–GOI (N-terminal GFP tag). Modules chosen from groups I, VII, VIII, IV, V and VI are connected in this order. The adh1 promoter (Padh1, group I) drives expression of the fusion gene GFP–GOI (groups VII and VIII). The plasmid digested with FseI can be targeted to the co2 locus of chromosome I by the co2 module (IV). Representative restriction sites are also shown. (b) List of module plasmids used for Golden Gate reaction to construct integration plasmids to express a GOI with an N-terminal FP tag (also see table 1). The GOI (cd) is a PCR product containing cohesive ends (‘c’ and ‘d’). To make control plasmids expressing fluorescent proteins without a GOI, a module from group VIIa can be used instead of VII and VIII. See figure 2b for other module groups. a–g: cohesive ends, sequences of which are shown in figure 2c.