TABLE 2.
Oligonucleotide primers used in this study
| Primer | Key feature | Sequence (5′–3′) |
|---|---|---|
| BLD1 | CaCas9 forward | ATCTCATTAGATTTGGAACTTGTGGGTT |
| BLD2 | CaCas9 reverse | TTCGAGCGTCCCAAAACCTTCT |
| BLD3 | SNR52 forward | AAGAAAGAAAGAAAACCAGGAGTGAA |
| BLD4 | sgRNA reverse | ACAAATATTTAAACTCGGGACCTGG |
| BLD5 | SNR52 NGG | GCGGCCGCAAGTGATTAGACT |
| BLD6 | sgRNA NGG | GCAGCTCAGTGATTAAGAGTAAAGATGG |
| BLD17 | CDA2 FLP forward | CGGTTTAATAGTCATTTAATAAAAACTCTTGAAATTCTTATCAAATAAACTAATCATTCTTCAATTACCATAAAGGGAACAAAAGCTGGG |
| BLD18 | CDA2 FLP reverse | CAACACTAAATTCTTCTTTGTAACCACCTACCTACCTACATACATACATACATACAATACAAGAATTTTTGTATTGATCTCTAGAACTAGTGGATCTG |
| BLD19 | DIT1 SNR52 reverse | GATGATTTACATGGAAAGGCCAAATTAAAAATAGTTTACGCAAGTC |
| BLD20 | DIT1 sgRNA forward | GCCTTTCCATGTAAATCATCGTTTTAGAGCTAGAAATAGCAAGTTAAA |
| BLD23 | DIT1 FLP forward | CGTTGAATTCAAATACAAGTAGTAATACCACGGTTGATACAGATTCGTTTGAACAAAAGCAACAACAAATATTGAAGCTAAAGGGAACAAAAGCTGGG |
| BLD24 | DIT1 FLP reverse | CGTTTTCACTCTCGTCACAGTTGGCCACAACCTATCGTCAGAAGAAGAAACAATAATCCAACGGAACAAACCTCTAGAACTAGTGGATCTG |
| BLD25 | DIT1 upstream verification forward | GGCTGCAATTTCCCCAAAAG |
| BLD26 | DIT1 downstream verification reverse | GCCAGAGTAGCCAACAAGTTA |
| BLD27 | CDA2 upstream verification forward | TTCCGGTGGTAATTTTGATGAGA |
| BLD29 | DIT1 midgene verification reverse | GGTCCCATGATGATGACAGG |
| BLD30 | CDA2 midgene verification reverse | TTTGTTGAGAGCATCCCACC |
| BLD31 | CDA2 downstream verification forward | GACTCGGTGCAATCTTGTCA |
| BLD36 | MUM3 sgRNA forward | GTAGTCCAAATATTTACTTCGTTTTAGAGCTAGAAATAGCAAGTTAAA |
| BLD37 | MUM3 SNR52 reverse | GAAGTAAATATTTGGACTACCAAATTAAAAATAGTTTACGCAAGTC |
| BLD38 | MUM3 FLP forward | GGCGACACTACCGATGCCAATCCCGCTGTGGTAGTAAGTAACCATGCATCTTTAGCGGACTGCTTTGTTATTCTAAAGGGAACAAAAGCTGGG |
| BLD39 | MUM3 FLP reverse | CTACCGGATTCAAAGAGATGAAAGTAGTAAATCAAGAATTTATAGTTTACCTATAGGTAGGATTCAAGAGAACCTCTAGAACTAGTGGATCTG |
| BLD40 | MUM3 upstream verification forward | CAGCATTTGAATAAGGTAAA |
| BLD41 | MUM3 midgene verification reverse | TGTCCCTGTAACGTTGCTCC |
| BLD42 | MUM3 downstream verification reverse | GGGAGATAGGTTTACTGATC |
| BLD43 | RRT8 sgRNA forward | GGTACGGAGTCGTTGCACTTGTTTTAGAGCTAGAAATAGCAAGTTAAA |
| BLD44 | RRT8 SNR52 reverse | AAGTGCAACGACTCCGTACCCAAATTAAAAATAGTTTACGCAAGTC |
| BLD45 | RRT8 FLP forward | CCAATCTTCTAGACGTGGGCTAAAGGCACATGCAAGATACTTTAAGTTGAAAGGGTTTCTGCGTAGCGACTAAAGGGAACAAAAGCTGGG |
| BLD46 | RRT8 FLP reverse | GCAGGTTGGTTGTTGAGGTCTAAGTTTAGTAGCAGCAATGAAGGTGGAGTTGCTGCTGGGTTTGGATGTGCTCTAGAACTAGTGGATCTG |
| BLD47 | RRT8 upstream verification forward | GTGGGCCCAATCATTGTCTTG |
| BLD48 | RRT8 midgene verification reverse | TGATAAATGGGAACAGCTCG |
| BLD49 | RRT8 downstream verification reverse | CGGGTGAAATCTTGACCAAC |
| BLD50 | SRT1 sgRNA forward | TGGGAAAGAACCTCGTGTCCGTTTTAGAGCTAGAAATAGCAAGTTAAA |
| BLD51 | SRT1 SNR52 reverse | GGACACGAGGTTCTTTCCCACAAATTAAAAATAGTTTACGCAAGTC |
| BLD52 | SRT1 FLP forward | CAAAATAAGTTAACCAGAAAAGCAATACTTGTCTTGTAAGTCGGAAAGCTTTTTACAAGATCATAGTTCCAGTAAAGGGAACAAAAGCTGGG |
| BLD53 | SRT1 FLP reverse | GAATCTATTCATGACAACTTTGCATATTCTAGCTAAAATACAAAATACAATCGTAAAGCAAGGCTCTAGAACTAGTGGATCTG |
| BLD54 | SRT1 upstream verification forward | GGATTAATTGTCGAGTGGCA |
| BLD55 | SRT1 midgene verification reverse | GTAATACTGGTGGAATAAC |
| BLD56 | SRT1 downstream verification reverse | TAAATAACCAGGTAGACTTG |
| BLD64 | CHS3 sgRNA forward | AAGGTGGACGTGAAGTTTATGTTTTAGAGCTAGAAATAGCAAGTTAAA |
| BLD65 | CHS3 SNR52 reverse | ATAAACTTCACGTCCACCTTCAAATTAAAAATAGTTTACGCAAGTC |
| BLD66 | CHS3 FLP forward | CCCTTGCATTAACACCAAAACTTATAGACAACAGAAACATTAGTCTTTTTTGTTTTCTACATTTATTCCTCTAAAGGGAACAAAAGCTGGG |
| BLD67 | CHS3 FLP reverse | GTACAATGCATGCAATAAACAAGGCAGAAATTTGAAATATTCTGGAGCCTCTATGTTATAAAGCAGCGTTGCTCTAGAACTAGTGGATCTG |
| BLD68 | CHS3 upstream verification forward | GTTTTCAATTACAATTAATC |
| BLD69 | CHS3 midgene verification reverse | CATAATCGTTAATTTCATCG |
| BLD70 | CHS3 downstream verification reverse | TTTGTGTTTGTAAGAGATTC |
| BLD71 | CDA2 sgRNA forward | ATCCGATCCATTTATTATGGGTTTTAGAGCTAGAAATAGCAAGTTAAA |
| BLD72 | CDA2 SNR52 reverse | CCATAATAAATGGATCGGATCAAATTAAAAATAGTTTACGCAAGTC |
| BLD73 | CDA2 verification reverse | CATGAATTTAGATTGAAGTC |
| BLD74 | DIT2 sgRNA forward | TTAGTGCTCATGGAGAATTGGTTTTAGAGCTAGAAATAGCAAGTTAAA |
| BLD75 | DIT2 SNR52 reverse | CAATTCTCCATGAGCACTAACAAATTAAAAATAGTTTACGCAAGTC |
| BLD76 | DIT2 FLP forward | GCACAGATAACCCTTTTGCTATTTGAGAACCATCCGGGTGATACTAGCCTTGCTCTTTCCTCTTAAACAAGTAAAGGGAACAAAAGCTGGG |
| BLD77 | DIT2 FLP reverse | GTGAGTGTGGGGTGTTTTCTGTTAGCAAACGCAAGTTATATACTATATGGTATGTACTGCATTCTTCATTCCTCTAGAACTAGTGGATCTG |
| BLD78 | DIT2 upstream verification forward | GACAATGAAATTTCCAAGACTCC |
| BLD79 | DIT2 midgene verification reverse | GGGCAACAACATCTCGGTATAG |
| BLD80 | DIT2 downstream verification reverse | AAATGCTTAGCTTACAGGGG |
| BLD97 | CIp10_CHS3 forward | CGATACCGTCGACCTCGAGGACAGACAGAGAGAGAGATCAGAGATTGAA |
| BLD104 | CIp10_CDA2 forward | CACTATAGGGCGAATTGGGTACCCGAAATTTAAAGAGACAATTGAAAAAATTACAAGGAG |
| BLD105 | CIp10_CDA2 reverse | GGGAACAAAAGCTGGGTACCTCATTTTGGGAAAGTTTTAATATAATCAATACCACC |
| BLD111 | CIp10_CHS3 reverse | CAAAAGCTGGGTACCGGGCCCTCAACCAGACCCCGAAGATGATCC |
| BLD112 | CIp10_MUM3 forward | CTTATCGATACCGTCGACCTCGAGATGGAATTCATTGAGCATTTAGGAGTCAAGC |
| BLD113 | CIp10_MUM3 reverse | CAAAAGCTGGGTACCGGGCCCCTACAGAGCTACAGAAAAATCATCTTGCAATATACG |
| BLD116 | CIp10_SRT1 forward | TACCGTCGACCTCGAGACAATTATAAATGTTTTCATTAGTGTTGGTAGTGTATCATATGC |
| BLD117 | CIp10_SRT1 reverse | GGGAACAAAAGCTGGGTACCGGGCCCTTAAATAACTGATGTAGCAGGTGGAGGG |
| BLD118 | CIp10_SRT1 verification | GGACAATCTCTTGTTTTTACC |
| BLD121 | CIp10 first half forward | CCCGGTACCCAGCTTTTGTTCCCTTTAGTG |
| BLD123 | CIp10 second half reverse | CTCGAGGTCGACGGTATCG |
| BLD125 | CIp10_RRT8 forward | CGATACCGTCGACCTCGAGATTGTTAATGGGACCACTAGGGGTG |
| BLD126 | CIp10_RRT8 reverse | CAAAAGCTGGGTACCGGGCCCTCAGATGGTATTTGTAGCAGTCTTTGGG |
| BLD142 | yEmRFP forward | ATGGTTTCAAAAGGTGAAGAAGATAATATGGC |
| BLD143 | CIp10_CDA2_yEmRFP reverse | TCTTCACCTTTTGAAACCATTTTTGGGAAAGTTTTAATATAATCAATACCACCAACAC |
| BLD144 | CIp10_MUM3_yEmRFP reverse | CTTCTTCACCTTTTGAAACCATCAGAGCTACAGAAAAATCATCTTGCAATATACG |
| BLD145 | CIp10_RRT8_yEmRFP reverse | CTTCTTCACCTTTTGAAACCATGATGGTATTTGTAGCAGTCTTTGGGG |
| BLD146 | CIp10_SRT1_yEmRFP reverse | CTTCTTCACCTTTTGAAACCATAATAACTGATGTAGCAGGTGGAGGG |
| BLD148 | yEmRFP reverse | CGATACCGTCGACCTCGAGTTATTTATATAATTCATCCATACCACCAGTTGAATGTCT |
| BLD153 | CIp10_RRT8_yEmRFP verification | TGTTACGACAAAAGGCTCAA |
| BLD154 | CIp10_CDA2_SAT1 verification | TACATTTATATAAAACCAGT |
| BLD155 | CIp10_CDA2_yEmRFP verification | GATGAAAAATAATAAAGGTT |
| BLD156 | CIp10_MUM3_yEmRFP verification | ACCGGTAGATCTGTTGATCA |
| BLD157 | CIp10_SRT1_yEmRFP verification | GGAGTTATTATAGAACTATT |
| OKZ67 | CIp10 first half reverse | GTATTCAACATTTCCGTGTCG |
| OKZ68 | CIp10 second half forward | CGACACGGAAATGTTGAATAC |