Table 1.
TRPY1 Plasmids Generated in the Laboratories of the Authors and Used in This Study
| Plasmid | TRPY1 Construct |
|---|---|
| Backbone: Yeast single-copy centromeric pRS316 vector | |
| pGS2062 | TRPY1 wild-type |
| pSB276 | TRPY1D398A |
| pSB277 | TRPY1D401A |
| pSB278 | TRPY1D405A |
| pSB279 | TRPY1D425A |
| pSB280 | TRPY1E428A,E429A |
| Backbone: Bicistronic eukaryotic expression (HEK-293 cells) | |
| pCAGGS-YVC1-IRES-GFP | TRPY1 wild-type |
| pCAGGS-YVC1D398A-IRES-GFP | TRPY1D398A |
| pCAGGS-YVC1D401A-IRES-GFP | TRPY1D401A |
| pCAGGS-YVC1D405A-IRES-GFP | TRPY1D405A |
| pCAGGS-YVC1D425A-IRES-GFP | TRPY1D425A |
| pCAGGS-YVC1E428A,E429A-IRES-GFP | TRPY1E428A,E429A |
The YVC1 gene encoding wild-type TRPY1 (NCBI accession number NM_001183506.1) was subcloned into the single-copy centromeric pRS316 vector (Sikorski and Hieter, 1989) under control of its own promoter and termination sequence. For some transformations (Figures S4A–S4C) the latter sequences were omitted. The eukaryotic expression vector pCAGGS has been described (Wissenbach et al., 2001).