TABLE 1.
Yeast plasmids
| Plasmida | Characteristics | Reference |
|---|---|---|
| YCpSc · U | A 0.5-kb DNA fragment bearing the yeast TOP1 promoter (pTOP1) was PCR amplified from yeast genomic DNA and ligated into HindIII-BamHI sites of pRS416b | This study |
| YCpSctop1T722A · U | A 2.9-kb BamHI-XbaI fragment excised from YCpGAL1top1T722A (23) was ligated into YCpSc · U | This study |
| YCpGPD · U | The 0.7-kb yeast GPD promoter, PCR amplified from yeast genomic DNA, was ligated into HindIII-BamHI sites of pRS416b | This study |
| YCpGPDyUBC9 · U | A 0.6-kb fragment, PCR amplified from YCpUBC9 · U (15), was ligated into the BamHI-NotI sites of YCpGPD · Ub | This study |
| YCpSc · H | The TOP1 promoter, excised from YCpSc · U, was cloned into HindIII-BamHI sites of pRS413 | This study |
| YCpSctop1T722A · H | top1T722A HIS3 CEN6 ARSH4; a 3.5-kb XhoI-NotI fragment, excised from YCpSctop1722A · U, was ligated into the same sites in pRS413 | This study |
| YCpUBC9 · L | A 2.2-kb BamHI-NotI genomic fragment from YCpUBC9 · U was ligated into pRS415 | This study |
| YCpGPD · L | The GPD promoter, excised from YCpGPD · U, was ligated into HindIII-BamHI sites of pRS415 | This study |
| YCpGPDhUBC9 · L | Human UBC9 cDNA sequences, PCR amplified from pooled human cDNA and cloned into pGEX4T3, were PCR amplified and ligated into the BamHI-NotI sites of YCpGPD · Lb | This study |
| YCpGPDhUBC9myc · L | A C-terminal Myc-tagged hUbc9, generated by PCR followed by cloning into BamHI-NotI sites of YCpGPD · Lb | This study |
| YCpGAL1 · U | A 0.8-kb fragment containing the yeast GAL1 promoter (pGAL1) was PCR amplified from yeast genomic DNA and inserted into the HindIII-BamHI sites of pRS416 | This study |
| YCpGAL1hUBC9 · U | hUBC9 cDNA ligated into the BamHI-NotI sites of YCpGAL1 · U | This study |
| YEp24-PL | Modified YEp24 vector with multiple cloning site of pBluescript II | 34 |
| YEpSIZ1 · U | A 4.3-kb SmaI-SacI genomic fragment containing SIZ1 was excised from a YEp-FY250 genomic DNA library clone (C. S. Lancaster and M.-A. Bjornsti unpublished results) and inserted into YEp24-PL | This study |
| YEpSIZ1 · T, · L, · U | The SmaI-SacI fragment from YEpSIZ1 · U inserted into pRS424, pRS425, or pRS426 | This study |
| YEpSIZ2 · L, · U | A 2.6-kb XhoI-NotI genomic fragment containing SIZ2 plus 500 bp 5′ of ATG was PCR amplified from genomic DNA, and cloned into pRS425 or pRS426, and confirmed by sequencingb | This study |
| YEpUBC9 · L | The BamHI-NotI genomic fragment from YCpUBC9 inserted into pRS425 | This study |
| YEpGAL1 · L | pGAL1 inserted into the HindIII-BamHI sites of pRS425 | This study |
| YEpGAL1yUBC9 · L | A yUBC9 PCR product was inserted into BamHI-NotI sites of YEpGAL1 · Lb | This study |
| YEpGAL1yUBC9myc · L | A C-terminal Myc epitope-tagged yUBC9 construct was PCR amplified and inserted into the BamHI-NotI sites of YEpGAL1 · Lb | This study |
| YEpGAL1hUBC9 · L | hUBC9 cDNA sequences ligated into BamHI-NotI sites of YEpGAL1 · L | This study |
| YEpGAL1hUBC9myc · L | A hUBC9myc PCR product was ligated into BamHI-NotI sites of YEpGAL1 · L | This study |
| YEpGAL1hUBC9flag · L | A C-terminal Flag-tagged hUBC9 construct was PCR amplified and inserted into the BamHI-NotI sites of YEpGAL1 · Lb | This study |
| YEpGAL1hubc9P123L · L | A Pro123(CCA)-to-Leu(CTA) mutation was generated by site-directed mutagenesis of YEpGAL1hUBC9 · L | This study |
a
A U, L, H, or T suffix indicates a URA3, LEU2, HIS3, or TRP1 marker, respectively.
b
Primer sequences are available upon request.