Table 2.
Primers and other oligonucleotides used in this study
| Name | Sequence (5′ → 3′)a,b |
|---|---|
| Primers for cloning | |
| pUC_AtKO_F | gcTCTAGAaggaggattacaaaatggccttcttctccatga |
| pUC_AtKO_R | ataagaatGCGGCCGCttaagaacgccttggattga |
| pUC_AtCPR2_F | gcTCTAGAaggaggattacaaaatgtcctcttcttcttcttc |
| pUC_AtCPR2_R | ataagaatGCGGCCGCttaccatacatctctaagatatc |
| pUC_CYP714A2_F | gcTCTAGAaggaggattacaaaatggagagcctggtggtc |
| pUC_CYP714A2_R | tcccCCCGGGttagacgacacggatcacga |
| pUC_trAtKO_F | gcTCTAGAaggaggattacaaaatggctttacttctggcagtttttaagaaacttctctccttctc |
| pUC_trCYP714A2_F | gcTCTAGAaggaggattacaaaatggctttacttctggcagtttttcgtgcggttgtcgagcag |
| pET_AtKO_F | cgGGATCCcatggccttcttctccatga |
| pET_AtKO_R | ccgCTCGAGagaacgccttggattgataat |
| pET_trAtKO_F | cgGGATCCatggctttacttctggcagt |
| Primers for Gibson assembly | |
| gibson_pV_F | ggccgctgcggtattttc |
| gibson_pV_GGPPS_R | ttccctacctccttttctttctcttcgctcacaattccacacaa |
| gibson_pV_CDPS_R | aatgttacctcctttgtttctgtttcgctcacaattccacacaa |
| gibson_pV_KS_R | tggtgggcctcctttaggtctcaaacgctcacaattccacacaa |
| gibson_pV_trAtKO_R | gttaatacctcctttttgtttggttcgctcacaattccacacaa |
| gibson_pV_trCYP714A2_R | gatgtaacctcctttatattcccctcgctcacaattccacacaa |
| gibson_GGPPS_F | tgagcgaagagaaagaaaaggaggtagggaaatggcgtttgaacagcgg |
| gibson_CDPS_F | tgagcgaaacagaaacaaaggaggtaacattatgaagaccggcttcatct |
| gibson_KS_F | tgagcgtttgagacctaaaggaggcccaccaatgaatctttcactatgcatc |
| gibson_trAtKO_F | tgagcgaaccaaacaaaaaggaggtattaacatggctttacttctggcagt |
| gibson_trCYP714A2_F | tgagcaggggaatataaaggaggttacatcatggctttacttctggcag |
| gibson_insert_R | aataccgcacagatgcgtaa |
| gibson_Fusion5_AtCPR2_F | ggtggcggcggaagcaggagatccggttctgg |
| gibson_Fusion5_trCYP714A2_R | cggatctcctgcttccgccgccaccgacgacacggatcacgac |
| gibson_Fusion10_AtCPR2_F | gtggcggtagtggcggtggtggaagtaggagatccggttctgg |
| gibson_Fusion10_trCYP714A2_R | cttccaccaccgccactaccgccaccaccgacgacacggatcacgac |
| gibson_Fusion15_AtCPR2_F | caggtggtgggggatctggtggcggtggcagtaggagatccggttctgg |
| gibson_Fusion15_trCYP714A2_R | ccgccaccagatcccccaccacctgacccccctcctccgacgacacggatcacgac |
| Primers for subcloning | |
| pSTVM2-sub-USER-3-F | agacagucataagtgcgg |
| pSTVM2-sub-USER-1-R | atgcaacucgtaggacag |
| pUC- sub-USER-3-F | agacagucaatctgctctgatgcc |
| pUC- sub-USER-1-R | atgcaacuca taatgaatcggccaac |
| pUC-sub-USER-1-F | agttgcaucccgactggaaagcg |
| pUC-sub-USER-2-F | atccatgucccgactggaaagcg |
| pUC-sub-USER-5-F | atatgcgaucccgactggaaagcg |
| pUC-sub-USER-2-R | acatggauatgcggtgtgaaatacc |
| pUC-sub-USER-5-R | atcgcatauatgcggtgtgaaataccg |
| pUC-sub-USER-3-R | actgtcuatgcggtgtgaaataccg |
| Primers for genome editing | |
| ldhA_up_F | aaacctttacgcgtaatgcg |
| ldhA_up_R | ctttccagtcgtgctataaacggcgagtt |
| GGPPS_F | tttatagcacgactggaaagcgggcag |
| GGPPS_R | gcaagattaaagaaaataccgcagcggcc |
| ldhA_Down_F | ggtattttctttaatcttgccgctcccc |
| ldhA_Down_R | ggttagcgcacatcatacg |
| malT_up_F | aaaaatggccgttgcgtatt |
| malT_up_R | tttccagtcgggacatggatagttaatcacttcactgtgga |
| CDPS_F | atccatgtcccgactggaaagcg |
| CDPS_R | tcatattacaatctcgaacacc |
| KS_F | tgttcgagattgtaatatgatttgagacctaaaggaggc |
| KS_R | actgtctatgcggtgtgaaataccg |
| malT_Down_F | tttcacaccgcatagacagtcaattgctgaagatgatggg |
| malT_Down_R | gccgggtaataccgtctc |
| Confirm_GGPPS_F | cggattgaagcggcaatg |
| Confirm _GGPPS_R | tgcccagcgtctcggcat |
| Confirm _CDPS/KS_F | aactcatcctcaataccaac |
| Confirm _CDPS/KS_R | ggctacccatgctgtgtc |
| Confirm _gdhA_F | ttatggctttacgcgccgc |
| Confirm _gdhA_R | gggacaattgaagaagaact |
| Oligonucleotides for deletion of gdhA | |
| gdhA-MAGE | aatatataagggttttatatctatggatcagacatattctctggcgcagggtgtgatttaagttgtaaatgcctgatggc |
aCapital letters indicate restriction sites
bUnderlining denotes 5′-UTR sequences