Development of fluorescent reporter tagged RIB gene cassettes for replicative transformation, early expression, and enhanced riboflavin production in Eremothecium ashbyi

► Reporter tagged shuttle vectors were constructed to initiate genetic engineering in E. ashbyi. ► Functionality of S. cerevisiae ARS elements were confirmed by reporter gene expression studies. ► Reporter genes mCherry and yEGFP, were expressed for the first time in E. ashbyi. ► RIB1 transformants produced 2.44 times more riboflavin than untransformed.