| Plasmid . | Description . | Reference . |
|---|---|---|
| pRS315 | CEN, LEU2 | Sikorski and Hieter, 1989 |
| pRS313 | CEN, HIS3 | Sikorski and Hieter, 1989 |
| pLP29 | pRS313; CDC12-GFP | Lippincott and Li, 1998 |
| pSB1 / pJT1520 | CEN, URA3, CDC11 | Versele et al., 2004 |
| pMVB39 / pJT1622 | CEN, URA3, CDC12 | Versele and Thorner, 2004 |
| pSC193 | URA3, prGAL1/10-NDT80-HA | Chu and Herskowitz, 1993 |
| YCpH-cdc12-6a | pLP29; cdc12-6 | This study |
| pGCF1 | pRS313; prCDC11-CDC11-ADH1(t)-CaURA3 | This study |
| pGCF2 | pRS315; prCDC12-CDC12-ADH1(t)-SpHIS5 | This study |
| pGCF3 | pRS315; prCDC11-SPR28-GFP-ADH1(t)-HygR | This study |
| pGCF4 | pRS313; prCDC12-SPR3-mCherry-ADH1(t)-KanR | This study |
| p3.8 | CDC3, CDC10, His6-SPR3 in E. coli | This study |
| p4.7 | CDC3, CDC10, His6-SPR3, SPR28 in E. coli | This study |
| p4.10 | CDC3, CDC10, His6-SPR3, CDC11 in E. coli | This study |
| pJT2485 | p406-GFP-2XPHOsh2 | Roy and Levine, 2004 |
| pML113 | CEN, LEU2, CDC12-eCitrine::his5MX | Nagaraj et al., 2008 |
| Plasmid . | Description . | Reference . |
|---|---|---|
| pRS315 | CEN, LEU2 | Sikorski and Hieter, 1989 |
| pRS313 | CEN, HIS3 | Sikorski and Hieter, 1989 |
| pLP29 | pRS313; CDC12-GFP | Lippincott and Li, 1998 |
| pSB1 / pJT1520 | CEN, URA3, CDC11 | Versele et al., 2004 |
| pMVB39 / pJT1622 | CEN, URA3, CDC12 | Versele and Thorner, 2004 |
| pSC193 | URA3, prGAL1/10-NDT80-HA | Chu and Herskowitz, 1993 |
| YCpH-cdc12-6a | pLP29; cdc12-6 | This study |
| pGCF1 | pRS313; prCDC11-CDC11-ADH1(t)-CaURA3 | This study |
| pGCF2 | pRS315; prCDC12-CDC12-ADH1(t)-SpHIS5 | This study |
| pGCF3 | pRS315; prCDC11-SPR28-GFP-ADH1(t)-HygR | This study |
| pGCF4 | pRS313; prCDC12-SPR3-mCherry-ADH1(t)-KanR | This study |
| p3.8 | CDC3, CDC10, His6-SPR3 in E. coli | This study |
| p4.7 | CDC3, CDC10, His6-SPR3, SPR28 in E. coli | This study |
| p4.10 | CDC3, CDC10, His6-SPR3, CDC11 in E. coli | This study |
| pJT2485 | p406-GFP-2XPHOsh2 | Roy and Levine, 2004 |
| pML113 | CEN, LEU2, CDC12-eCitrine::his5MX | Nagaraj et al., 2008 |
The cdc12-6 allele was first introduced into a CDC12-GFP gene on pLP29 by digestion with BfuAI and co-transformation of the cut plasmid into YMVB61 along with the PCR-amplified cdc12-6 gene from DDY1462. Plasmids from transformants that displayed temperature sensitivity upon loss of the URA3-marked CDC12 plasmid were rescued to E. coli and sequenced to confirm the presence of the cdc12-6 allele (K391N L392stop).