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Additional file 11: Figure S1. Attempts to increase flux towards cytosolic FPP by CrFPPS accumulation. Different orientations with the mCerulean3 reporter and co-expression of AgBS and corresponding bisabolene yields (fg cell−1) are depicted (constructs indicated on the left). Error bars represent 95% confidence intervals. Figure S2. Detection of full-length protein products of 3 independent cell lines transformed with various constructs. Each cell line is denoted by individual numbers, constructs are specified at the top, and corresponding bisabolene yields are depicted in Fig. 2. Total cellular proteins were run on a 10% SDS PAGE gel and subjected to Western blotting with α-GFP antibody. M marker (PageRuler prestained protein ladder, ThermoFisher Scientific). WT parental strain UVM4. P positive control expressing AgBS-mVenus, AgBS-mCerulean3, and AgBS-mRuby2 to the cytosol. Positive control and ERG20 protein products show previously observed pattern of protein breakage in the reporter [7]. Predicted protein sizes are: ~ 124.0 kDa for AgBS-mVenus, ~ 70.2 kDa for ERG20-mCerulean3, and ~ 67.7 kDa for mCerulean3-ERG20 which lacks a StrepII tag on its C terminus. Figure S3. Detection of full-length protein products of 3 independent cell lines transformed with various constructs. Each cell line is denoted by individual numbers, constructs are specified at the top, and corresponding bisabolene yields are depicted in Fig. 3. Total cellular proteins were run on a 10% SDS PAGE gel and subjected to Western blotting with α-GFP antibody. M marker (PageRuler prestained protein plus ladder, ThermoFisher Scientific). WT parental strain UVM4. Y mVenus expressed to the cytosol. Predicted protein sizes are: ~ 124.0 kDa for AgBS-mVenus, ~ 161.9 kDa for AgBS-mVenus-ERG20. Ponceau S stain is shown as loading control on the nitrocellulose membrane. Signal for construct vi, strain #3 was below detection limit, but bisabolene could be detected in GC–MS measurements, indicating full-length expression. Figure S4. Detection of full-length protein products of 3 independent cell lines transformed with various constructs. Each cell line is denoted by individual numbers, constructs are specified at the top, and corresponding bisabolene yields are depicted in Fig. 4. Total cellular proteins were run on a 10% SDS PAGE gel and subjected to Western blotting with α-GFP and α-StrepII antibodies. M marker (PageRuler prestained protein ladder, ThermoFisher Scientific). WT parental strain UVM4. P positive control expressing AgBS-mVenus, AgBS-mCerulean3, and AgBS-mRuby2 to the cytosol. Positive control and ERG20 protein products show previously observed pattern of protein breakage in the reporter [7]. Predicted protein sizes are: ~ 124.0 kDa for AgBS-mVenus, ~ 123.1 kDa for AgBS-mCerulean3, ~ 120.9 kDa for AgBS-mRuby2, ~ 123.7 kDa for AgBS-aadA, ~ 67.7 kDa for mCerulean3-ERG20 which lacks a StrepII tag on its C terminus, ~ 161.9 kDa for AgBS-mVenus-ERG20, and ~ 162.2 kDa for AgBS-mCerulean3-ERG20. Figure S5. Detection of full-length protein products of 3 independent cell lines transformed with various constructs. Each cell line is denoted by individual numbers, constructs are specified at the top, and corresponding bisabolene yields are depicted in Fig. 3 and Additional file 11 Fig. S1. Total cellular proteins were run on a 10% SDS PAGE gel and subjected to Western blotting with α-GFP antibodies. M marker (PageRuler prestained protein ladder, ThermoFisher Scientific). WT parental strain UVM4. CrFPPS protein products show previously observed pattern of protein breakage in the reporter [7]. Predicted protein sizes are: ~ 160.9 kDa for AgBS-mVenus-CrFPPS, ~ 124.0 kDa for AgBS-mVenus, ~ 68.5 kDa for CrFPPS-mCerulean3, and ~ 67.0 kDa for mCerulean3-CrFPPS which lacks a StrepII tag on its C terminus. Figure S6. Calibration line and correspondent equation for bisabolene quantification. GC/MS peak areas were normalized to the internal standard peak area, and plotted against the bisabolene concentration in dodecane.

Additional file 1: Table S1. Summary table of S. bombicola���s predicted transporters. Table S2. Keys for TransAAP substrate prediction, grouped. Table S3. S. bombicola strains used in this research. Table S4. Primers used in this research. Table S5. Maximum specific growth rate (��) of selected S. bombicola ABC transporter knockouts cultivated on SD medium. Table S6. List of compounds used for disc diffusion assay, their concentration in DMSO and supplier. Figure S1. Disc diffusion assay with polyoxyethylene-9-lauryl ether of selected S. bombicola ABC transporter deletion mutants compared to wild type. Figure S2. Disc diffusion assay of S. bombicola ��SbCdr4 and S. bombicola ��SbYor1.2 compared to wild type. The concentration of the applied medium chain alkanes and fatty alcohols is given in Table S6. Figure S3. Comparison of growth among different SL transporter deletion strains on shake flask scale by OD600 and pH measurements every 24 h (a) without the addition of rapeseed oil and (b) with the addition of 37.5 g/L rapeseed oil.