Additional file 1: Figure S1. Testing tRNATrp-UCA variants for temperature-dependent behaviour. Figure S2. Growth curve of C. reinhardtii cell lines, showing that the temperature-sensitive tRNA (tCI) does not cause a growth defect. Figure S3. Induction of CrCD protein in the C. reinhardtii CD/2* + tCI cell line at low temperatures. Figure S4. Growth curves for C. reinhardtii CD/4* + tCI following induction at 15–25 °C. Figure S5. PCR confirming homoplasmic transgene integration into C. reinhardtii TN72 using transformation plates incubated at 30 °C. Figure S6. REVERT total protein staining of E. coli western blot membrane in Fig. 5e, confirming equal loading and blotting across lanes. Figure S7. Investigation of readthrough effect of four drugs as an alternative method of induction. Figure S8. Minimum free energy calculations for each tRNA variant at 15–35 °C. Figure S9. Detection of tryptophan in purified CrCD protein from C. reinhardtii CD (positive control) and CD/6* + tCI cell lines. Table S1. Primers used to alter trnWUCA to make variants 1 to 4. Table S2. Conditions for C. reinhardtii induction experiments. Sequences 1 to 9. Includes DNA sequences for tRNAs, genes CD and TPS4, 16S promoter and plasmids pWUCA3 and pWUCA4.