Fig. 5: Reconstruction of transcription units.
From: Building a eukaryotic chromosome arm by de novo design and synthesis
a Strategy to identify functional synthetic promoters and terminators to support both essential and nonessential genes. eCDS, CDS of essential gene. neCDS, CDS of nonessential gene. URA3-WT, the CEN plasmid containing wild-type essential gene and URA3. b Functional analysis of reconstructed genes by tetrad analysis. The corresponding heterozygous diploid containing a reconstructed TU (rTU) was sporulated and dissected. One representative tetrad was shown for each strain. For YLL035W, the ATG codon was mutated (marked with *) to avoid impact on the expression of YLL036W. Red circle, the spores containing disrupted native gene and reconstructed TU. c Functional complementation test of reconstructed YLL002W and YLL006W. WT, BY4741. + and − indicate strains with or without the reconstructed TU. d Comparison of sequence identity in CDS between the 24 rTUs and corresponding genes in Sc2.0 project. The bounds of the box were the upper and lower quartile with the median value in the center. The whiskers indicated the minimum and maximum. e Length comparison of Pro and Ter between the 24 rTUs and the native ones used in ptWT25. The data are presented as the mean and SD. Unpaired t test (two-tail) was used for comparison in (d) (p = 3.57E-11) and (e) (p = 2.15E-06 for Pro, p = 2.77E-05 for Ter). Source data are provided as a Source Data file.
