TABLE 4.
List of primers used in the study
| Primer no. |
Primer sequence (5′–3′)a | Purpose |
|---|---|---|
| BP196 | aggcaatagcatttccatctggtttcttgtcgaccatatgGGAACATCTGAAATTGGTTC | Primer to amplify ORI410 to clone in pGEM-CaHIS1 |
| BP197 | gaatactcaagctatgcatccaacgcgttgggagctctccTTGATGATTGGATCGGGTTC | Primer to amplify ORI410 to clone in pGEM-CaHIS1 |
| BP1266 | gcatgctcccggccgccatggccgcgggatGTAACGGCCGCCAGTGTG | Primer to amplify CdARG4 and CmLEU2 for cloning in pGEM and pGEM-ORI410 |
| BP1267 | catccaacgcgttgggagctctcccatatgCCAGTGTGATGGATATCTGCAG | Primer to amplify CdARG4 and CmLEU2 for cloning in pGEM and pGEM-ORI410 |
| BP1262 | CATATGGGAGAGCTCCCAACGCGTTG | Forward primer to amplify pGEM backbone from pGEM-CaHIS1 to clone CdARG4 and CmLEU2 |
| BP1265 | CATATGGGAACATCTGAAATTGGTTCTTTGGTAGATCTGCC | Forward primer to amplify pGEM-ORI410 backbone from pGEM-CaHIS1-ORI410 to clone CdARG4 and CmLEU2 |
| BP1263 | ATCCCGCGGCCATGGCGG | Reverse primer to amplify pGEM backbone from pGEM-CaHIS1 and pGEM-CaHIS1-ORI410 to clone CdARG4 and CmLEU2 |
| BP1246 | GTCGACCTGCAGGCGGCC | Primer to amplify pGEM-CaURA3 to clone ORI |
| BP1247 | GGAGAGCTCCCAACGCGTTG | Primer to amplify pGEM-CaURA3 to clone ORI |
| BP1248 | aatcactagtgcggccgcctgcaggtcgacTTGTAGATTTCAAAAATGCTTC | Primer to clone ORI7-L1 in pGEM-CaURA3 |
| BP1249 | gctatgcatccaacgcgttgggagctctccGATTTGTGTGTGCTTACTAGAG | Primer to clone ORI7-L1 in pGEM-CaURA3 |
| BP1250 | aatcactagtgcggccgcctgcaggtcgacTTGTGTAGTAAAGGGTTGTTG | Primer to clone ORI7-R1 in pGEM-CaURA3 |
| BP1251 | gctatgcatccaacgcgttgggagctctccAGTTAGGAAGAGTATAAATATGTGTAGTC | Primer to clone ORI7-R1 in pGEM-CaURA3 |
| BP1198 | ttctgcagatatccatcacactggcatatgACAAAAAATCATTAGCAAAATATTC | Primer to amplify ORI410_178 to clone in pGEM-CmLEU2 |
| BP1199 | gctatgcatccaacgcgttgggagctctccCCAGTGGAATTTGCAACC | Primer to amplify ORI410_178 to clone in pGEM-CmLEU2 |
| BP1200 | ttctgcagatatccatcacactggcatatgACTTTCAGAAATTGGTTGG | Primer to amplify ORI410_97 to clone in pGEM-CmLEU2 |
| BP1201 | gctatgcatccaacgcgttgggagctctccACACAAAAAATCATTAGCAAAATATTC | Primer to amplify ORI410_97 to clone in pGEM-CmLEU2 |
| BP1214 | ttctgcagatatccatcacactggcatatgAGCAGTTTTAAAATAAATAGGG | Primer to amplify proORI1088 to clone in pGEM-CmLEU2 |
| BP1215 | gctatgcatccaacgcgttgggagctctccTTGGATTATCAAAAAATCATTAG | Primer to amplify proORI1088 to clone in pGEM-CmLEU2 |
| BP1194 | ttctgcagatatccatcacactggcatatgTGTTGCAAAATATGAGTAAAAAAA | Primer to amplify ORI246 to clone in pGEM-CmLEU2 |
| BP1195 | gctatgcatccaacgcgttgggagctctccACAACGGAGGGTAAGGTG | Primer to amplify ORI246 to clone in pGEM-CmLEU2 |
| BP1192 | ttctgcagatatccatcacactggcatatgTGGTTATGTACTTGATCACCC | Primer to amplify ORI1055 to clone in pGEM-CmLEU2 |
| BP1193 | gctatgcatccaacgcgttgggagctctccTACAGAATGAGTAATATACAATGTTTG | Primer to amplify ORI1055 to clone in pGEM-CmLEU2 |
| BP1196 | ttctgcagatatccatcacactggcatatgATATATTTGTGATTCAACCACAC | Primer to amplify ORI1046 to clone in pGEM-CmLEU2 |
| BP1197 | gctatgcatccaacgcgttgggagctctccCAAAAATATCTCGTGAATCTTTTC | Primer to amplify ORI1046 to clone in pGEM-CmLEU2 |
| BP1186 | ttctgcagatatccatcacactggcatatgCACATGTTAAAATAGTGAAGGAG | Primer to amplify ARS1max to clone in pGEM-CmLEU2 |
| BP1187 | gctatgcatccaacgcgttgggagctctccAAAGCTTACATTTTATGTTAGCTG | Primer to amplify ARS1max to clone in pGEM-CmLEU2 |
| BP1188 | ttctgcagatatccatcacactggcatatgTCAACATCTTTGGATAATATCAG | Primer to amplify panARS to clone in pGEM-CmLEU2 |
| BP1189 | gctatgcatccaacgcgttgggagctctccTAGTGCTGATTATGATTTGACG | Primer to amplify panARS to clone in pGEM-CmLEU2 |
| BP1179 | CATATGCCAGTGTGATGGATATCTG | Primer to amplify pGEM-CmLEU2 to clone ORIs |
| BP1180 | GGAGAGCTCCCAACGCGT | Primer to amplify pGEM-CmLEU2 to clone ORIs |
| BP1204 | ACTGGCCGTCGTTTTACA | Primer to amplify linear plasmids without TEL |
| BP1205 | GAATTGTAATACGACTCACTATAGGG | Primer to amplify linear plasmids without TEL |
| BP1252 | CCGTACACCAAGAAGTTAGACATCCGTACACCAActtaagggatccgcatgctcccggccgccatg | Primer to amplify linear plasmids with 1.5× TEL repeat |
| BP1253 | CCGTACACCAAGAAGTTAGACATCCGTACACCAActtaagggatccgggcccaattcgccctatag | Primer to amplify linear plasmids with 1.5× TEL repeat |
| BP1254 | CCGTACACCAAGAAGTTAGACATCCGTACACCAAGAAGTTAGACATCCGTACACCAA
cttaagggatccgcatgctcccggccgccatg |
Primer to amplify linear plasmids with 2.5× TEL repeats |
| BP1255 | CCGTACACCAAGAAGTTAGACATCCGTACACCAAGAAGTTAGACATCCGTACACCAA
cttaagggatccgggcccaattcgccctatag |
Primer to amplify linear plasmids with 2.5× TEL repeats |
| BP1843 | CAAGGCGAGTTACATGATCC | Primer to amplify AMPR for qPCR |
| BP1844 | GGATGGCATGACAGTAAGAG | Primer to amplify AMPR for qPCR |
| BP285 | TTTGTACTTAGCGGCTACCTG | Primer to amplify chromosome 1L CEN for qPCR |
| BP317 | GAAAGAAGTGGGAGGAAAGGG | Primer to amplify chromosome 1L CEN for qPCR |
| BP1869 | CATGTATGGTAATCCAAATGGG | Forward primer that anneals outside 5′ UTR of CaHIS1 |
| BP1870 | AACACGGTGCACCAGTC | Reverse primer that anneals outside 3′ UTR of CaHIS1 |
| BP1841 | GGCTGGCTGGTTTATTGC | Reverse primer that anneals to AMPR gene |
| BP1873 | GGTAATGTAATGGACGAATTGAAG | Forward primer that anneals within CaHIS1 |
| BP1857 | CAACCTGGGTATTGATATGTTG | Reverse primer that anneals to CmLEU2 promoter |
a
Sequences in lowercase indicate regions homologous to the plasmid.