Overview Data points present in this dataset were obtained following the protocol described in dx.doi.org/10.17504/protocols.io.kfnctme. We picked transformed colonies and cultured in 400 μL TAP medium for 7 days in Deep-well plates (Corning Axygen®, No.: PDW500CS, Thermo Fisher Scientific Inc., Waltham, MA), covered with Breathe-Easy® (Sigma-Aldrich®). Cultivation was performed on a rotary shaker, set to 150 rpm, under constant illumination (50 μmol photons/m2s). Then 100 μL sample were transferred clear bottom 96-well plate (Corning Costar, Tewksbury, MA, USA) and fluorescence was measured using an Infinite® M200 PRO plate reader (Tecan, Männedorf, Switzerland). Supernatant samples were obtained by spinning Deep-well plates at 3000 × g for 10 min and transferring 100 μL from each well to the clear bottom 96-well plate (Corning Costar, Tewksbury, MA, USA), followed by fluorescence measurement. To compare the constructs, R Statistic version 3.3.3 was used to perform one-way ANOVA (with Tukey's test), and to test statistical hypotheses, the significance level was set at 0.05. Graphs were generated in RStudio v1.0.136. The codes are deposit herein. Info ANOVA_Turkey_Sub.R -> code for ANOVA analysis in R statistic 3.3.3 Anova_Output_Summary_Guide.pdf -> Explain the ANOVA files content Analysis_Raw_FP.xlsx -> File with raw values organized in a spreadsheet pRFU_FLUORESCENT PROTEIN_+_bk.csv -> relative supernatant mCherry fluorescence dataset of positive colonies, blanked with parental wild-type cc1690 cell of Chlamydomonas reinhardtii sup_RFU_FLUORESCENT PROTEIN_+_bk.csv -> supernatant mCherry fluorescence dataset of positive colonies, blanked with parental wild-type cc1690 cell of Chlamydomonas reinhardtii who_RFU_FLUORESCENT PROTEIN_+_bk.csv -> whole culture mCherry fluorescence dataset of positive colonies, blanked with parental wild-type cc1690 cell of Chlamydomonas reinhardtii pRFU_FLUORESCENT PROTEIN_+_bk.doc -> ANOVA of relative supernatant mCherry fluorescence dataset of positive colonies, blanked with parental wild-type cc1690 cell of Chlamydomonas reinhardtii sup_RFU_FLUORESCENT PROTEIN_+_bk.doc -> ANOVA of supernatant mCherry fluorescence dataset of positive colonies, blanked with parental wild-type cc1690 cell of Chlamydomonas reinhardtii who_RFU_FLUORESCENT PROTEIN_+_bk.doc -> ANOVA of whole culture mCherry fluorescence dataset of positive colonies, blanked with parental wild-type cc1690 cell of Chlamydomonas reinhardtii Consider citing our work. 1. Molino JVD, de Carvalho JCM, Mayfield S. Evaluation of secretion reporters to microalgae biotechnology: blue to red fluorescent proteins. Algal Res. 2018;31: 252–261. doi:10.1016/j.algal.2018.02.018