Supplementary Figure 3: Burden imposed by the four synthetic constructs.

(a) Transcription profiles of the synthetic constructs pD864-LacZ, pLys-M1 and pSB1C3-Lux in DH10B cells 15 (top) and 60 (bottom) min post-induction. Traces show the normalized reads mapping to the plasmid constructs. Solid grey regions show average, and transparent red regions span from minimum to maximum of transcription profiles generated from three biological replicates. Genetic designs below each profile are drawn using SBOLv notation and show the precise location of each genetic part. For pD864-LacZ, only reads uniquely mapping to the plasmid lacZ gene (and not the chromosomal copy) are shown. These are used to estimate the level of plasmid lacZ expression (blue shaded regions, see Online Methods section). (b) Transcriptional induction of the four constructs represented by mean FPKM values in MG1655 at 1h post induction. Green bars are data for the calculated GFP FPKM and colour intensity relates to the time point; grey bars are data for the FPKM corresponding to the construct gene; * denotes empty plasmid controls. Dots and bars show individual measurements and their mean, respectively. Error bars show the standard deviation of three independent repeats. (c) Percentage of available capacity 1h post induction. In the left panels, plots show GFP mRNA FPKM in construct samples compared to the pD864 control. In the right panels, plots show the same comparison but using GFP fluorescence production rate data from the plate-based burden assay (Figure 1c). Dots show single repeats while bars show the mean value. Error bars show standard deviation of three independent repeats.