Fig. 4
Promoters to construct the Shikimate pathway refactoring library. a Transcriptional strengths of constitutive promoters from K. marxianus and S. cerevisiae were quantified by overexpression of EGFP on a low copy-number plasmid. Three characterized promoters from K. marxianus CBS 6556 (PKmTEF3, PKmPGK, and PKmTDH3) were used to construct the Shikimate pathway library. b The temperature effect on chromosomal expression of EGFP from variable strength promoters. Expression cassettes of EGFP driven by PKmTEF3, PKmPGK and PKmTDH3 were integrated into the genome of K. marxianus CBS 6556 ura3Δ his3Δ at the ABZ1 locus. All fluorescence measurements were taken using flow cytometry. All experiments were performed in biological triplicates. Bars represent the mean of background-subtracted fluorescence, while error bars represent the standard deviation