Table 1. Plasmids used in the present study.
(a), Dieter Oesterhelt (Max Planck Institute for Biochemistry, Martinsreid, Germany); (b), [10]; (c), Béatrice Vallée; (Center for Molecular Biophysics, CNRS, Orleans, France) (d), the present study.
| Plasmid number | Plasmids | Inserts | Source |
|---|---|---|---|
| T1118 | pESC-URA | None | (a) |
| 1119 | pSTS30a | LAG1 (N-term., c-Myc-tagged) | (a) |
| 1121 | pSTS30b | LAC1 (N-term., FLAG-tagged) | (a) |
| 1548 | pSTS30c | LAG1 (N-term., FLAG-tagged) | (b) |
| 1582 | Full-length of SUC2 | (c) | |
| 1750 | pRS414-PT | Promoter and terminator of LAG1 | (d) |
| 1751 | pRS414-LAG1 | LAG1 (N-term., FLAG-tagged) | (d) |
| 1752 | pRS416-PT | Promoter and terminator of LAG1 | (d) |
| 1753 | pRS416-LAG1 | LAG1 (N-term., Myc-tagged) | (d) |
| 1754 | pSUC2A | SUC2 reporter cassette A | (d) |
| 1755–1765 | pESC-LAG1-N, 1–8, C | SacI site is introduced into various regions of LAG1 | (d) |
| 1766–1776 | pESC-LAC1-N, 1–8, C | SacI site is introduced into various regions of LAC1 | (d) |
| 1777–1787 | pESC-LAG1-SUC2A N, 1–8, C | SUC2A cassette is inserted into SacI sites of no. 1755–1765 | (d) |
| 1788–1798 | pESC-LAC1-SUC2A N, 1–8, C | SUC2A cassette is inserted into SacI sites of no. 1755–1765 | (d) |
| 1799–1809 | pRS414-LAG1-SUC2A N, 1–8, C | LAG1–SUC2A fusion constructs expressed under LAG1’s promoter | (d) |
| 1810 | pESC-LAG1-fXa4 | fXa cleavage site is inserted in Pro245 of LAG1 | (d) |
| 1811 | pESC-LAG1-fXa5 | fXa cleavage site is inserted in Thr272 of LAG1 | (d) |
| 1812 | pESC-LAG1-fXa6 | fXa cleavage site is inserted in Leu298 of LAG1 | (d) |
| 1813 | pESC-LAG1-fXa7 | fXa cleavage site is inserted in Phe333 of LAG1 | (d) |
| 1814 | pESC-LAG1-fXa8 | fXa cleavage site is inserted in Gly386 of LAG1 | (d) |
| 1815 | pESC-LAC1-fXa4 | fXa cleavage site is inserted in Pro245 of LAC1 | (d) |
| 1816 | pESC-LAC1-fXa5 | fXa cleavage site is inserted in Thr272 of LAC1 | (d) |
| 1817 | pESC-LAC1-fXa6 | fXa cleavage site is inserted in Leu298 of LAC1 | (d) |
| 1818 | pESC-LAC1-fXa7 | fXa cleavage site is inserted in Phe333 of LAC1 | (d) |
| 1819 | pESC-LAC1-fXa8 | fXa cleavage site is inserted in Gly386 of LAC1 | (d) |
| 1820–1824 | pRS414-LAG1-fXa4–8 | LAG1–fXa-cleavage-site fusion constructs expressed under LAG1's promoter | (d) |
| 1825 | pRS414-lag1(H255,256A) | lag1 (H255A, H256A) | (d) |
| 1826 | pRS414-lag1(L262A) | lag1 (L262A) | (d) |
| 1827 | pRS414-lag1(S266A) | lag1 (S266A) | (d) |
| 1828 | pRS414-lag1(D283A) | lag1 (D283A) | (d) |
| 1829 | pRS414-lag1(D283,286A) | lag1 (D283A, D286A) | (d) |
| 1853 | pESC-LAG1-SUC2A-4H | H255A H256A mutations were introduced into no. 1781 | (d) |
| 1854 | pESC-LAG1-SUC2A-5H | H255A H256A mutations were introduced into no. 1782 | (d) |
| 1855 | pESC-LAG1-SUC2A-6H | H255A H256A mutations were introduced into no. 1784 | (d) |
| 1856 | pESC-LAG1-SUC2A-7H | H255A H256A mutations were introduced into no. 1785 | (d) |
| 1857 | pESC-LAG1-SUC2A-4D | D283A D286A mutations were introduced into no. 1781 | (d) |
| 1858 | pESC-LAG1-SUC2A-5D | D283A D286A mutations were introduced into no. 1782 | (d) |
| 1859 | pESC-LAG1-SUC2A-6D | D283A D286A mutations were introduced into no. 1784 | (d) |
| 1860 | pESC-LAG1-SUC2A-7D | D283A D286A mutations were introduced into no. 1785 | (d) |