Table 1.
Plasmids used in this study
| Name (enzyme)a | Vector | Insert |
|---|---|---|
| pB-KRE5-HA (SacI) | pBluescript II KS(+)b | KRE5(+3627c–+4095)-3HA-HDEL-stop-TRP1d,e-KRE5(+4115–+4720) |
| pB-HA-KRE6 (SacI) | pBluescript II KS(+) | KRE6(−1275–−712)-CgTRP1f-KRE6(−639–−1)-ATG-3HA-KRE6(+1–+1486) |
| pB-BIG1-HA (NotI/XhoI) | pBluescript II KS(+) | BIG1(−605–+1092)-3HA-stop-TRP1d-BIG1(+1111–+2082) |
| pB-sec12-URA3 (XbaI/XhoI) | pBluescript II KS(+) | sec12-4(−783–+1967)g-URA3h-SEC12(+1965–+2340) |
| pRS314-ATG22-HA | pRS314i (TRP1, CEN) | ATG22(−869–+1584)-3HA-stop-ADH1td,j |
| pB-DRS2-HA (KpnI/SacI) | pBluescript II KS(+) | DRS2(+3224–+4065)-3HA-stop-TRP1d-DRS2(+3657–+4065) |
| pRS424-DRS2-HA | pRS424i (TRP1, 2 μ) | DRS2(−617–+4065)-3HA-stop-ADH1t |
| pT10-GUP1-HA | pGCT10e (TRP1, CEN) | GUP1(−857–+1680)-3HA-stop-ADH1t |
| pT20-GUP1-HA | pGCT20e (TRP1, 2 μ) | GUP1(−857–+1680)-3HA-stop-ADH1t |
| pRS314-HA-OPI3 | pRS314 (TRP1, CEN) | OPI3(−643–−1)-ATG-3HA-OPI3(+1–+853) |
| pB-PMR1-HA (BamI/XhoI) | pBluescript II KS(+) | PMR1(+2046–+2850)-2HA-stop-TRP1d-PMR1(+2277–+2850) |
| pYEX-ROT1-HA-His | pYEX4Tk (leu2d, URA3, 2 μ) | TDH3(−1037–−7)-ROT1-(+1–+693)-HA-His8-stop-TDH3t(+1967–+2107) |
| pYEX-rot1-2-HA-His | pYEX4T (leu2d, URA3, 2 μ) | TDH3(−1037–−7)-rot1-2-(+1–+693)-HA-His8-stop-TDH3t(+1967–+2107) |
| pB-kar2-1-LEU2 (SpeI/SalI) | pBluescript II KS(+) | kar2-1(+1161–+2643)-LEU2-KAR2(+2080–+2990) |
a The plasmid was digested with indicated enzyme(s) and introduced to yeast cells.
b Stratagene, La Jolla, CA.
c The A residue of the initiation codon was set at +1.
d Derived from pGCT10.
f Candida glabrata TRP1; derived from pUC18-CgTRP1 (provided by Dr. Mukai, Osaka University).
g The sec12-4 mutant gene was obtained from the genome of strain MBY10-7A (Nakano et al., 1988) by PCR.
h Derived from YCp50 (Rose et al., 1987).
j Terminator.
k Clontech Lab, Mountain View, CA.