Authentic samples of many carotenoids are commercially available, but are often too expensive and of insufficient purity. As an alternative, a wide range of carotenoids can be biosynthesized in recombinant Escherichia coli strains. E. coli cells naturally produce the carotenoid biosynthetic precursors and provide a blank slate for manufacturing different target carotenoids, depending on the introduced genes encoding carotenoid biosynthetic enzymes. This chapter presents a series of plasmids that carry genes encoding carotenoid biosynthetic enzymes and methods to produce in E. coli 21 different carotenoids and two apocarotenoids, which can be used as authentic standards. For the structural confirmation of these carotenoids or apocarotenoids, we show their individual spectral data containing MS/MS and UV-visible, along with their biosynthetic routes in the E. coli transformants.