TABLE 2.
Plasmids used in this study
| Plasmid | Descriptiona | Reference or source |
|---|---|---|
| pRS414 | CEN6 ARS1 TRP1 | 49 |
| pRS416 | CEN6 ARS1 URA3 | 49 |
| pTEB16 | CEN ARS1 TRP1 GAL3 | 8 |
| pGP15b | CEN ARS1 URA3 GAL80GFP | This study |
| pGP15Δ | CEN ARS1 URA3 GAL80 | This study |
| pGP31 | CEN ARS1 URA3 NLS-GAL80GFP | This study |
| pGP31Δ | CEN ARS1 URA3 NLS-GAL80 | This study |
| pGP31m | CEN ARS1 URA3 mNLS-GAL80GFP | This study |
| pGP31mΔ | CEN ARS1 URA3 mNLS-GAL80 | This study |
| pGP13 | 2μm LEU2 PADH1-GAL80GFP | This study |
| pGP20c | CEN ARS1 TRP NLS-GAL3 | This study |
| pGP20GFP | CEN ARS1 TRP NLS-GAL3GFP | This study |
| pGP25 | CEN ARS1 URA3 PADH2-GAL80GFP | This study |
| pGP25-S0 | CEN ARS1 URA3 PADH2-GAL80S0-GFP | This study |
| pGP25-S1 | CEN ARS1 URA3 PADH2-GAL80S1-GFP | This study |
| pGP25-S2 | CEN ARS1 URA3 PADH2-GAL80S2-GFP | This study |
| pGP26 | CEN ARS1 TRP PADH2-NLS-GAL3 | This study |
| pGP53 | CEN ARS1 URA3 PGAL-CYC-NLS-GAL80GFP | This study |
| pGP10 | XhoI-GFP in pUC19 | This study |
a
PADH1 and PADH2 designate the ADH1 and the ADH2 promoters, respectively. PGAL-CYC designates the hybrid GAL-CYC promoter. Native promoters were used in all other constructs.
b
pGP15 has a unique AatII site at the N terminus of Gal80p that allows insertion of NLS or mNLS oligonucleotides; GFP was inserted at a unique NarI site that was previously created at the Gal80p C terminus.
c
NLS was inserted in pGP20 at a unique AatII site that was previously created at the N terminus of Gal3p.