Appendix 1—key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Escherichia coli) | ER2566 | New England Biolabs | Cat# C2566I | Chemically competent cells |
| Strain, strain background (Escherichia coli) | ER2566 ∆folA/∆thyA (–Lon) | Reynolds et al. Cell 2011 | Chemically competent and electrocompetent cells | |
| Strain, strain background (Escherichia coli) | ER2566 ∆folA/∆thyA (+Lon) | This work | Chemically competent and electrocompetent cells | |
| Recombinant DNA reagent | SMT101 (plasmid) | This work | Dual expression of DHFR and TYMS, in vivo assays, chloramphenicol (35 µg/mL final concentration) | |
| Recombinant DNA reagent | SMT201 (plasmid) | This work | SMT101 with TET promter for TYMS, in vivo assays, Chloramphenicol (35 µg/mL final concentration) | |
| Recombinant DNA reagent | SMT205 (plasmid) | This work | SMT201 with mutated RBS for DHFR, in vivo assays, Chloramphenicol (35 µg/mL final concentration) | |
| Recombinant DNA reagent | SMT215 (plasmid) | This work | SMT205 with DHFR-FLAG-tag, western blot, Chloramphenicol (35 µg/mL final concentration) | |
| Recombinant DNA reagent | KR101/SMT301 (plasmid) | Reynolds et al. Cell 2011 | His8-tag, Heterologous expression, NiNTA purfication, kanamycin (50 µg/mL final concentration) |
|
| Recombinant DNA reagent | pSIM6 (plasmid) | Blomfield et al., 1991 | Lambda Red recombinase expression, temperature-sensitive promoter, ampicillin/carbenicilin (100 µg/mL final concentration) |
|
| Recombinant DNA reagent | pIB279 (plasmid) | Blomfield et al., 1991 | KAN-SacB cassette for positive/negative selection, ampicillin/carbenicilin (100 µg/mL final concentration) | |
| Sequence-based reagent | TetDuet1_sense | This work | Mutagenic PCR primer | ccgCTTAAGtcgaacagaaagtaatcgtattgtacatccctatc |
| Sequence-based reagent | TetDuet2_anti | This work | Mutagenic PCR primer | gatagggatgtcaatctctatcactgatagggatgtacaatacg |
| Sequence-based reagent | TetDuet3_sense | This work | Mutagenic PCR primer | agagattgacatccctatcagtgatagagatactgagcacatcag |
| Sequence-based reagent | TetDuet4_anti | This work | Mutagenic PCR primer | ctttaatgaattcggtcagtgcgtcctgctgatgtgctcagtatctc |
| Sequence-based reagent | TetDuet5_sense | This work | Mutagenic PCR primer | cactgaccgaattcattaaagaggagaaaggtaccatatggc |
| Sequence-based reagent | TetDuet_5flanking | This work | Mutagenic PCR primer | ccgcttaagtcgaacagaaag |
| Sequence-based reagent | TetDuet_3flanking | This work | Mutagenic PCR primer | cggagatctgccatatggtacc |
| Sequence-based reagent | WT_DHFR_pos2_fwd | This work | Mutagenic PCR primer | NNSAGTCTGATTGCGGCGTTAG |
| Sequence-based reagent | WT_DHFR_pos2_fwd2 | This work | Mutagenic PCR primer | NNSAGTCTGATTGCGGCGTTAG |
| Sequence-based reagent | WT_DHFR_pos3_fwd | This work | Mutagenic PCR primer | NNSCTGATTGCGGCGTTAGCG |
| Sequence-based reagent | WT_DHFR_pos4_fwd | This work | Mutagenic PCR primer | NNSATTGCGGCGTTAGCGGTA |
| Sequence-based reagent | WT_DHFR_pos5_fwd | This work | Mutagenic PCR primer | NNSGCGGCGTTAGCGGTAGAT |
| Sequence-based reagent | WT_DHFR_pos6_fwd | This work | Mutagenic PCR primer | NNSGCGTTAGCGGTAGATCGC |
| Sequence-based reagent | WT_DHFR_pos7_fwd | This work | Mutagenic PCR primer | NNSTTAGCGGTAGATCGCGTTATC |
| Sequence-based reagent | WT_DHFR_pos8_fwd | This work | Mutagenic PCR primer | NNSGCGGTAGATCGCGTTATCG |
| Sequence-based reagent | WT_DHFR_pos8_fwd2 | This work | Mutagenic PCR primer | NNSGCGGTAGATCGCGTTATCG |
| Sequence-based reagent | WT_DHFR_pos9_fwd | This work | Mutagenic PCR primer | NNSGTAGATCGCGTTATCGGCATG |
| Sequence-based reagent | WT_DHFR_pos10_fwd | This work | Mutagenic PCR primer | NNSGATCGCGTTATCGGCATGG |
| Sequence-based reagent | WT_DHFR_pos11_fwd | This work | Mutagenic PCR primer | NNSCGCGTTATCGGCATGGAAAA |
| Sequence-based reagent | WT_DHFR_pos12_fwd | This work | Mutagenic PCR primer | NNSGTTATCGGCATGGAAAACGC |
| Sequence-based reagent | WT_DHFR_pos13_fwd | This work | Mutagenic PCR primer | NNSATCGGCATGGAAAACGCC |
| Sequence-based reagent | WT_DHFR_pos14_fwd | This work | Mutagenic PCR primer | NNSGGCATGGAAAACGCCATG |
| Sequence-based reagent | WT_DHFR_pos15_fwd | This work | Mutagenic PCR primer | NNSATGGAAAACGCCATGCCG |
| Sequence-based reagent | WT_DHFR_pos16_fwd | This work | Mutagenic PCR primer | NNSGAAAACGCCATGCCGTGG |
| Sequence-based reagent | WT_DHFR_pos17_fwd | This work | Mutagenic PCR primer | NNSAACGCCATGCCGTGGAAC |
| Sequence-based reagent | WT_DHFR_pos18_fwd | This work | Mutagenic PCR primer | NNSGCCATGCCGTGGAACCTG |
| Sequence-based reagent | WT_DHFR_pos19_fwd | This work | Mutagenic PCR primer | NNSATGCCGTGGAACCTGCCT |
| Sequence-based reagent | WT_DHFR_pos20_fwd | This work | Mutagenic PCR primer | NNSCCGTGGAACCTGCCTGCC |
| Sequence-based reagent | WT_DHFR_pos21_fwd | This work | Mutagenic PCR primer | NNSTGGAACCTGCCTGCCGAT |
| Sequence-based reagent | WT_DHFR_pos22_fwd | This work | Mutagenic PCR primer | NNSAACCTGCCTGCCGATCTC |
| Sequence-based reagent | WT_DHFR_pos22_fwd2 | This work | Mutagenic PCR primer | NNSAACCTGCCTGCCGATCTC |
| Sequence-based reagent | WT_DHFR_pos23_fwd | This work | Mutagenic PCR primer | NNSCTGCCTGCCGATCTCGCC |
| Sequence-based reagent | WT_DHFR_pos24_fwd | This work | Mutagenic PCR primer | NNSCCTGCCGATCTCGCCTGG |
| Sequence-based reagent | WT_DHFR_pos25_fwd | This work | Mutagenic PCR primer | NNSGCCGATCTCGCCTGGTTT |
| Sequence-based reagent | WT_DHFR_pos26_fwd | This work | Mutagenic PCR primer | NNSGATCTCGCCTGGTTTAAACGC |
| Sequence-based reagent | WT_DHFR_pos27_fwd | This work | Mutagenic PCR primer | NNSCTCGCCTGGTTTAAACGCAACA |
| Sequence-based reagent | WT_DHFR_pos28_fwd | This work | Mutagenic PCR primer | NNSGCCTGGTTTAAACGCAACAC |
| Sequence-based reagent | WT_DHFR_pos29_fwd | This work | Mutagenic PCR primer | NNSTGGTTTAAACGCAACACCTTAAATAAAC |
| Sequence-based reagent | WT_DHFR_pos30_fwd | This work | Mutagenic PCR primer | NNSTTTAAACGCAACACCTTAAATAAACCCG |
| Sequence-based reagent | WT_DHFR_pos31_fwd | This work | Mutagenic PCR primer | NNSAAACGCAACACCTTAAATAAACCCGTG |
| Sequence-based reagent | WT_DHFR_pos32_fwd | This work | Mutagenic PCR primer | NNSCGCAACACCTTAAATAAACCCGT |
| Sequence-based reagent | WT_DHFR_pos33_fwd | This work | Mutagenic PCR primer | NNSAACACCTTAAATAAACCCGTGATTATGG |
| Sequence-based reagent | WT_DHFR_pos34_fwd | This work | Mutagenic PCR primer | NNSACCTTAAATAAACCCGTGATTATGGG |
| Sequence-based reagent | WT_DHFR_pos35_fwd | This work | Mutagenic PCR primer | NNSTTAAATAAACCCGTGATTATGGGCC |
| Sequence-based reagent | WT_DHFR_pos36_fwd | This work | Mutagenic PCR primer | NNSAATAAACCCGTGATTATGGGCC |
| Sequence-based reagent | WT_DHFR_pos37_fwd | This work | Mutagenic PCR primer | NNSAAACCCGTGATTATGGGCC |
| Sequence-based reagent | WT_DHFR_pos38_fwd | This work | Mutagenic PCR primer | NNSCCCGTGATTATGGGCCGC |
| Sequence-based reagent | WT_DHFR_pos39_fwd | This work | Mutagenic PCR primer | NNSGTGATTATGGGCCGCCATAC |
| Sequence-based reagent | WT_DHFR_pos40_fwd | This work | Mutagenic PCR primer | NNSATTATGGGCCGCCATACCT |
| Sequence-based reagent | WT_DHFR_pos41_fwd | This work | Mutagenic PCR primer | NNSATGGGCCGCCATACCTGG |
| Sequence-based reagent | WT_DHFR_pos42_fwd | This work | Mutagenic PCR primer | NNSGGCCGCCATACCTGGGAA |
| Sequence-based reagent | WT_DHFR_pos42_fwd2 | This work | Mutagenic PCR primer | NNSGGCCGCCATACCTGGGAATC |
| Sequence-based reagent | WT_DHFR_pos43_fwd | This work | Mutagenic PCR primer | NNSCGCCATACCTGGGAATCAATC |
| Sequence-based reagent | WT_DHFR_pos43_fwd2 | This work | Mutagenic PCR primer | NNSCGCCATACCTGGGAATCAATC |
| Sequence-based reagent | WT_DHFR_pos44_fwd | This work | Mutagenic PCR primer | NNSCATACCTGGGAATCAATCGGTC |
| Sequence-based reagent | WT_DHFR_pos45_fwd | This work | Mutagenic PCR primer | NNSACCTGGGAATCAATCGGTC |
| Sequence-based reagent | WT_DHFR_pos46_fwd | This work | Mutagenic PCR primer | NNSTGGGAATCAATCGGTCGTC |
| Sequence-based reagent | WT_DHFR_pos47_fwd | This work | Mutagenic PCR primer | NNSGAATCAATCGGTCGTCCGTTG |
| Sequence-based reagent | WT_DHFR_pos48_fwd | This work | Mutagenic PCR primer | NNSTCAATCGGTCGTCCGTTGC |
| Sequence-based reagent | WT_DHFR_pos49_fwd | This work | Mutagenic PCR primer | NNSATCGGTCGTCCGTTGCCA |
| Sequence-based reagent | WT_DHFR_pos50_fwd | This work | Mutagenic PCR primer | NNSGGTCGTCCGTTGCCAGGAC |
| Sequence-based reagent | WT_DHFR_pos51_fwd | This work | Mutagenic PCR primer | NNSCGTCCGTTGCCAGGACGC |
| Sequence-based reagent | WT_DHFR_pos52_fwd | This work | Mutagenic PCR primer | NNSCCGTTGCCAGGACGCAAA |
| Sequence-based reagent | WT_DHFR_pos53_fwd | This work | Mutagenic PCR primer | NNSTTGCCAGGACGCAAAAATATTATCC |
| Sequence-based reagent | WT_DHFR_pos54_fwd | This work | Mutagenic PCR primer | NNSCCAGGACGCAAAAATATTATCCTCAG |
| Sequence-based reagent | WT_DHFR_pos55_fwd | This work | Mutagenic PCR primer | NNSGGACGCAAAAATATTATCCTCAGCAG |
| Sequence-based reagent | WT_DHFR_pos56_fwd | This work | Mutagenic PCR primer | NNSCGCAAAAATATTATCCTCAGCAGTCAA |
| Sequence-based reagent | WT_DHFR_pos57_fwd | This work | Mutagenic PCR primer | NNSAAAAATATTATCCTCAGCAGTCAACCGG |
| Sequence-based reagent | WT_DHFR_pos58_fwd | This work | Mutagenic PCR primer | NNSAATATTATCCTCAGCAGTCAACCGGGTA |
| Sequence-based reagent | WT_DHFR_pos59_fwd | This work | Mutagenic PCR primer | NNSATTATCCTCAGCAGTCAACCG |
| Sequence-based reagent | WT_DHFR_pos60_fwd | This work | Mutagenic PCR primer | NNSATCCTCAGCAGTCAACCG |
| Sequence-based reagent | WT_DHFR_pos61_fwd | This work | Mutagenic PCR primer | NNSCTCAGCAGTCAACCGGGT |
| Sequence-based reagent | WT_DHFR_pos62_fwd | This work | Mutagenic PCR primer | NNSAGCAGTCAACCGGGTACG |
| Sequence-based reagent | WT_DHFR_pos63_fwd | This work | Mutagenic PCR primer | NNSAGTCAACCGGGTACGGAC |
| Sequence-based reagent | WT_DHFR_pos64_fwd | This work | Mutagenic PCR primer | NNSCAACCGGGTACGGACGAT |
| Sequence-based reagent | WT_DHFR_pos65_fwd | This work | Mutagenic PCR primer | NNSCCGGGTACGGACGATCGC |
| Sequence-based reagent | WT_DHFR_pos66_fwd | This work | Mutagenic PCR primer | NNSGGTACGGACGATCGCGTA |
| Sequence-based reagent | WT_DHFR_pos66_fwd2 | This work | Mutagenic PCR primer | NNSGGTACGGACGATCGCGTAAC |
| Sequence-based reagent | WT_DHFR_pos67_fwd | This work | Mutagenic PCR primer | NNSACGGACGATCGCGTAACG |
| Sequence-based reagent | WT_DHFR_pos67_fwd2 | This work | Mutagenic PCR primer | NNSACGGACGATCGCGTAACG |
| Sequence-based reagent | WT_DHFR_pos68_fwd | This work | Mutagenic PCR primer | NNSGACGATCGCGTAACGTGG |
| Sequence-based reagent | WT_DHFR_pos69_fwd | This work | Mutagenic PCR primer | NNSGATCGCGTAACGTGGGTG |
| Sequence-based reagent | WT_DHFR_pos70_fwd | This work | Mutagenic PCR primer | NNSCGCGTAACGTGGGTGAAG |
| Sequence-based reagent | WT_DHFR_pos71_fwd | This work | Mutagenic PCR primer | NNSGTAACGTGGGTGAAGTCGG |
| Sequence-based reagent | WT_DHFR_pos72_fwd | This work | Mutagenic PCR primer | NNSACGTGGGTGAAGTCGGTG |
| Sequence-based reagent | WT_DHFR_pos73_fwd | This work | Mutagenic PCR primer | NNSTGGGTGAAGTCGGTGGAT |
| Sequence-based reagent | WT_DHFR_pos73_fwd2 | This work | Mutagenic PCR primer | NNSTGGGTGAAGTCGGTGGATG |
| Sequence-based reagent | WT_DHFR_pos74_fwd | This work | Mutagenic PCR primer | NNSGTGAAGTCGGTGGATGAAGC |
| Sequence-based reagent | WT_DHFR_pos74_fwd2 | This work | Mutagenic PCR primer | NNSGTGAAGTCGGTGGATGAAGC |
| Sequence-based reagent | WT_DHFR_pos75_fwd | This work | Mutagenic PCR primer | NNSAAGTCGGTGGATGAAGCCAT |
| Sequence-based reagent | WT_DHFR_pos76_fwd | This work | Mutagenic PCR primer | NNSTCGGTGGATGAAGCCATC |
| Sequence-based reagent | WT_DHFR_pos77_fwd | This work | Mutagenic PCR primer | NNSGTGGATGAAGCCATCGCG |
| Sequence-based reagent | WT_DHFR_pos78_fwd | This work | Mutagenic PCR primer | NNSGATGAAGCCATCGCGGCG |
| Sequence-based reagent | WT_DHFR_pos79_fwd | This work | Mutagenic PCR primer | NNSGAAGCCATCGCGGCGTGT |
| Sequence-based reagent | WT_DHFR_pos80_fwd | This work | Mutagenic PCR primer | NNSGCCATCGCGGCGTGTGGT |
| Sequence-based reagent | WT_DHFR_pos80_fwd2 | This work | Mutagenic PCR primer | NNSGCCATCGCGGCGTGTGG |
| Sequence-based reagent | WT_DHFR_pos81_fwd | This work | Mutagenic PCR primer | NNSATCGCGGCGTGTGGTGAC |
| Sequence-based reagent | WT_DHFR_pos82_fwd | This work | Mutagenic PCR primer | NNSGCGGCGTGTGGTGACGTA |
| Sequence-based reagent | WT_DHFR_pos82_fwd2 | This work | Mutagenic PCR primer | NNSGCGGCGTGTGGTGACGTACCAGAAATC |
| Sequence-based reagent | WT_DHFR_pos83_fwd | This work | Mutagenic PCR primer | NNSGCGTGTGGTGACGTACCA |
| Sequence-based reagent | WT_DHFR_pos84_fwd | This work | Mutagenic PCR primer | NNSTGTGGTGACGTACCAGAAATCAT |
| Sequence-based reagent | WT_DHFR_pos84_fwd2 | This work | Mutagenic PCR primer | NNSTGTGGTGACGTACCAGAAATCATG |
| Sequence-based reagent | WT_DHFR_pos85_fwd | This work | Mutagenic PCR primer | NNSGGTGACGTACCAGAAATCATGG |
| Sequence-based reagent | WT_DHFR_pos86_fwd | This work | Mutagenic PCR primer | NNSGACGTACCAGAAATCATGGTGATTGG |
| Sequence-based reagent | WT_DHFR_pos87_fwd | This work | Mutagenic PCR primer | NNSGTACCAGAAATCATGGTGATTGGCGG |
| Sequence-based reagent | WT_DHFR_pos88_fwd | This work | Mutagenic PCR primer | NNSCCAGAAATCATGGTGATTGGCGG |
| Sequence-based reagent | WT_DHFR_pos89_fwd | This work | Mutagenic PCR primer | NNSGAAATCATGGTGATTGGCGGCG |
| Sequence-based reagent | WT_DHFR_pos89_fwd2 | This work | Mutagenic PCR primer | NNSGAAATCATGGTGATTGGCGGC |
| Sequence-based reagent | WT_DHFR_pos90_fwd | This work | Mutagenic PCR primer | NNSATCATGGTGATTGGCGGC |
| Sequence-based reagent | WT_DHFR_pos91_fwd | This work | Mutagenic PCR primer | NNSATGGTGATTGGCGGCGGTC |
| Sequence-based reagent | WT_DHFR_pos92_fwd | This work | Mutagenic PCR primer | NNSGTGATTGGCGGCGGTCGC |
| Sequence-based reagent | WT_DHFR_pos93_fwd | This work | Mutagenic PCR primer | NNSATTGGCGGCGGTCGCGTTTA |
| Sequence-based reagent | WT_DHFR_pos94_fwd | This work | Mutagenic PCR primer | NNSGGCGGCGGTCGCGTTTAT |
| Sequence-based reagent | WT_DHFR_pos95_fwd | This work | Mutagenic PCR primer | NNSGGCGGTCGCGTTTATGAA |
| Sequence-based reagent | WT_DHFR_pos95_fwd2 | This work | Mutagenic PCR primer | NNSGGCGGTCGCGTTTATGAAC |
| Sequence-based reagent | WT_DHFR_pos96_fwd | This work | Mutagenic PCR primer | NNSGGTCGCGTTTATGAACAGTTCTT |
| Sequence-based reagent | WT_DHFR_pos97_fwd | This work | Mutagenic PCR primer | NNSCGCGTTTATGAACAGTTCTTGC |
| Sequence-based reagent | WT_DHFR_pos98_fwd | This work | Mutagenic PCR primer | NNSGTTTATGAACAGTTCTTGCCAAAAGCGC |
| Sequence-based reagent | WT_DHFR_pos99_fwd | This work | Mutagenic PCR primer | NNSTATGAACAGTTCTTGCCAAAAGCGC |
| Sequence-based reagent | WT_DHFR_pos100_fwd | This work | Mutagenic PCR primer | NNSGAACAGTTCTTGCCAAAAGCGCAAAAAC |
| Sequence-based reagent | WT_DHFR_pos101_fwd | This work | Mutagenic PCR primer | NNSCAGTTCTTGCCAAAAGCGCAAAAAC |
| Sequence-based reagent | WT_DHFR_pos102_fwd | This work | Mutagenic PCR primer | NNSTTCTTGCCAAAAGCGCAAAAAC |
| Sequence-based reagent | WT_DHFR_pos103_fwd | This work | Mutagenic PCR primer | NNSTTGCCAAAAGCGCAAAAACTGTAT |
| Sequence-based reagent | WT_DHFR_pos104_fwd | This work | Mutagenic PCR primer | NNSCCAAAAGCGCAAAAACTGTATCTGA |
| Sequence-based reagent | WT_DHFR_pos104_fwd2 | This work | Mutagenic PCR primer | NNSCCAAAAGCGCAAAAACTGTATCTG |
| Sequence-based reagent | WT_DHFR_pos105_fwd | This work | Mutagenic PCR primer | NNSAAAGCGCAAAAACTGTATCTGACG |
| Sequence-based reagent | WT_DHFR_pos106_fwd | This work | Mutagenic PCR primer | NNSGCGCAAAAACTGTATCTGACG |
| Sequence-based reagent | WT_DHFR_pos107_fwd | This work | Mutagenic PCR primer | NNSCAAAAACTGTATCTGACGCATATCGAC |
| Sequence-based reagent | WT_DHFR_pos107_fwd2 | This work | Mutagenic PCR primer | NNSCAAAAACTGTATCTGACGCATATCG |
| Sequence-based reagent | WT_DHFR_pos108_fwd | This work | Mutagenic PCR primer | NNSAAACTGTATCTGACGCATATCGAC |
| Sequence-based reagent | WT_DHFR_pos109_fwd | This work | Mutagenic PCR primer | NNSCTGTATCTGACGCATATCGACG |
| Sequence-based reagent | WT_DHFR_pos110_fwd | This work | Mutagenic PCR primer | NNSTATCTGACGCATATCGACGCA |
| Sequence-based reagent | WT_DHFR_pos111_fwd | This work | Mutagenic PCR primer | NNSCTGACGCATATCGACGCAG |
| Sequence-based reagent | WT_DHFR_pos112_fwd | This work | Mutagenic PCR primer | NNSACGCATATCGACGCAGAAGT |
| Sequence-based reagent | WT_DHFR_pos113_fwd | This work | Mutagenic PCR primer | NNSCATATCGACGCAGAAGTGGAAG |
| Sequence-based reagent | WT_DHFR_pos114_fwd | This work | Mutagenic PCR primer | NNSATCGACGCAGAAGTGGAAG |
| Sequence-based reagent | WT_DHFR_pos115_fwd | This work | Mutagenic PCR primer | NNSGACGCAGAAGTGGAAGGC |
| Sequence-based reagent | WT_DHFR_pos116_fwd | This work | Mutagenic PCR primer | NNSGCAGAAGTGGAAGGCGAC |
| Sequence-based reagent | WT_DHFR_pos117_fwd | This work | Mutagenic PCR primer | NNSGAAGTGGAAGGCGACACC |
| Sequence-based reagent | WT_DHFR_pos118_fwd | This work | Mutagenic PCR primer | NNSGTGGAAGGCGACACCCAT |
| Sequence-based reagent | WT_DHFR_pos118_fwd2 | This work | Mutagenic PCR primer | NNSGTGGAAGGCGACACCCATTTC |
| Sequence-based reagent | WT_DHFR_pos119_fwd | This work | Mutagenic PCR primer | NNSGAAGGCGACACCCATTTCC |
| Sequence-based reagent | WT_DHFR_pos120_fwd | This work | Mutagenic PCR primer | NNSGGCGACACCCATTTCCCG |
| Sequence-based reagent | WT_DHFR_pos121_fwd | This work | Mutagenic PCR primer | NNSGACACCCATTTCCCGGATTAC |
| Sequence-based reagent | WT_DHFR_pos122_fwd | This work | Mutagenic PCR primer | NNSACCCATTTCCCGGATTACGA |
| Sequence-based reagent | WT_DHFR_pos123_fwd | This work | Mutagenic PCR primer | NNSCATTTCCCGGATTACGAGCC |
| Sequence-based reagent | WT_DHFR_pos124_fwd | This work | Mutagenic PCR primer | NNSTTCCCGGATTACGAGCCG |
| Sequence-based reagent | WT_DHFR_pos125_fwd | This work | Mutagenic PCR primer | NNSCCGGATTACGAGCCGGAT |
| Sequence-based reagent | WT_DHFR_pos126_fwd | This work | Mutagenic PCR primer | NNSGATTACGAGCCGGATGACTG |
| Sequence-based reagent | WT_DHFR_pos127_fwd | This work | Mutagenic PCR primer | NNSTACGAGCCGGATGACTGG |
| Sequence-based reagent | WT_DHFR_pos128_fwd | This work | Mutagenic PCR primer | NNSGAGCCGGATGACTGGGAA |
| Sequence-based reagent | WT_DHFR_pos129_fwd | This work | Mutagenic PCR primer | NNSCCGGATGACTGGGAATCG |
| Sequence-based reagent | WT_DHFR_pos130_fwd | This work | Mutagenic PCR primer | NNSGATGACTGGGAATCGGTATTCAG |
| Sequence-based reagent | WT_DHFR_pos131_fwd | This work | Mutagenic PCR primer | NNSGACTGGGAATCGGTATTCAGC |
| Sequence-based reagent | WT_DHFR_pos131_fwd2 | This work | Mutagenic PCR primer | NNSGACTGGGAATCGGTATTCAGC |
| Sequence-based reagent | WT_DHFR_pos132_fwd | This work | Mutagenic PCR primer | NNSTGGGAATCGGTATTCAGCGAATT |
| Sequence-based reagent | WT_DHFR_pos133_fwd | This work | Mutagenic PCR primer | NNSGAATCGGTATTCAGCGAATTCCAC |
| Sequence-based reagent | WT_DHFR_pos134_fwd | This work | Mutagenic PCR primer | NNSTCGGTATTCAGCGAATTCCAC |
| Sequence-based reagent | WT_DHFR_pos135_fwd | This work | Mutagenic PCR primer | NNSGTATTCAGCGAATTCCACGATG |
| Sequence-based reagent | WT_DHFR_pos135_fwd2 | This work | Mutagenic PCR primer | NNSGTATTCAGCGAATTCCACGATGC |
| Sequence-based reagent | WT_DHFR_pos136_fwd | This work | Mutagenic PCR primer | NNSTTCAGCGAATTCCACGATGC |
| Sequence-based reagent | WT_DHFR_pos136_fwd2 | This work | Mutagenic PCR primer | NNSTTCAGCGAATTCCACGATGC |
| Sequence-based reagent | WT_DHFR_pos137_fwd | This work | Mutagenic PCR primer | NNSAGCGAATTCCACGATGCTG |
| Sequence-based reagent | WT_DHFR_pos138_fwd | This work | Mutagenic PCR primer | NNSGAATTCCACGATGCTGATGC |
| Sequence-based reagent | WT_DHFR_pos139_fwd | This work | Mutagenic PCR primer | NNSTTCCACGATGCTGATGCG |
| Sequence-based reagent | WT_DHFR_pos140_fwd | This work | Mutagenic PCR primer | NNSCACGATGCTGATGCGCAG |
| Sequence-based reagent | WT_DHFR_pos140_fwd2 | This work | Mutagenic PCR primer | NNSCACGATGCTGATGCGCAG |
| Sequence-based reagent | WT_DHFR_pos141_fwd | This work | Mutagenic PCR primer | NNSGATGCTGATGCGCAGAACT |
| Sequence-based reagent | WT_DHFR_pos142_fwd | This work | Mutagenic PCR primer | NNSGCTGATGCGCAGAACTCTC |
| Sequence-based reagent | WT_DHFR_pos143_fwd | This work | Mutagenic PCR primer | NNSGATGCGCAGAACTCTCACAG |
| Sequence-based reagent | WT_DHFR_pos144_fwd | This work | Mutagenic PCR primer | NNSGCGCAGAACTCTCACAGC |
| Sequence-based reagent | WT_DHFR_pos145_fwd | This work | Mutagenic PCR primer | NNSCAGAACTCTCACAGCTATTGCTTTG |
| Sequence-based reagent | WT_DHFR_pos146_fwd | This work | Mutagenic PCR primer | NNSAACTCTCACAGCTATTGCTTTGAGATT |
| Sequence-based reagent | WT_DHFR_pos147_fwd | This work | Mutagenic PCR primer | NNSTCTCACAGCTATTGCTTTGAGATTCT |
| Sequence-based reagent | WT_DHFR_pos148_fwd | This work | Mutagenic PCR primer | NNSCACAGCTATTGCTTTGAGATTCTGG |
| Sequence-based reagent | WT_DHFR_pos149_fwd | This work | Mutagenic PCR primer | NNSAGCTATTGCTTTGAGATTCTGGAG |
| Sequence-based reagent | WT_DHFR_pos150_fwd | This work | Mutagenic PCR primer | NNSTATTGCTTTGAGATTCTGGAGCG |
| Sequence-based reagent | WT_DHFR_pos151_fwd | This work | Mutagenic PCR primer | NNSTGCTTTGAGATTCTGGAGCG |
| Sequence-based reagent | WT_DHFR_pos152_fwd | This work | Mutagenic PCR primer | NNSTTTGAGATTCTGGAGCGGC |
| Sequence-based reagent | WT_DHFR_pos153_fwd | This work | Mutagenic PCR primer | NNSGAGATTCTGGAGCGGCGG |
| Sequence-based reagent | WT_DHFR_pos154_fwd | This work | Mutagenic PCR primer | NNSATTCTGGAGCGGCGGTAA |
| Sequence-based reagent | WT_DHFR_pos155_fwd | This work | Mutagenic PCR primer | NNSCTGGAGCGGCGGTAACAG |
| Sequence-based reagent | WT_DHFR_pos156_fwd | This work | Mutagenic PCR primer | NNSGAGCGGCGGTAACAGGCG |
| Sequence-based reagent | WT_DHFR_pos157_fwd | This work | Mutagenic PCR primer | NNSCGGCGGTAACAGGCGTCG |
| Sequence-based reagent | WT_DHFR_pos158_fwd | This work | Mutagenic PCR primer | NNSCGGTAACAGGCGTCGACA |
| Sequence-based reagent | WT_DHFR_pos159_fwd | This work | Mutagenic PCR primer | NNSTAACAGGCGTCGACAAGCT |
| Sequence-based reagent | WT_DHFR_pos2_rev | This work | Mutagenic PCR primer | CATGGTATATCTCCTTATTAAAGTTAAA |
| Sequence-based reagent | WT_DHFR_pos2_rev2 | This work | Mutagenic PCR primer | CATGGTATATCTCATTATTAAAGTTAAAC |
| Sequence-based reagent | WT_DHFR_pos3_rev | This work | Mutagenic PCR primer | GATCATGGTATATCTCCTTATTAAAGTT |
| Sequence-based reagent | WT_DHFR_pos4_rev | This work | Mutagenic PCR primer | ACTGATCATGGTATATCTCCTTATTAAA |
| Sequence-based reagent | WT_DHFR_pos5_rev | This work | Mutagenic PCR primer | CAGACTGATCATGGTATATCTCCTTATT |
| Sequence-based reagent | WT_DHFR_pos6_rev | This work | Mutagenic PCR primer | AATCAGACTGATCATGGTATATCTCCTT |
| Sequence-based reagent | WT_DHFR_pos7_rev | This work | Mutagenic PCR primer | CGCAATCAGACTGATCATGGTATATCT |
| Sequence-based reagent | WT_DHFR_pos8_rev | This work | Mutagenic PCR primer | CGCCGCAATCAGACTGATC |
| Sequence-based reagent | WT_DHFR_pos8_rev2 | This work | Mutagenic PCR primer | CGCCGCAATCAGACTGATC |
| Sequence-based reagent | WT_DHFR_pos9_rev | This work | Mutagenic PCR primer | TAACGCCGCAATCAGACTGA |
| Sequence-based reagent | WT_DHFR_pos10_rev | This work | Mutagenic PCR primer | CGCTAACGCCGCAATCAG |
| Sequence-based reagent | WT_DHFR_pos11_rev | This work | Mutagenic PCR primer | TACCGCTAACGCCGCAAT |
| Sequence-based reagent | WT_DHFR_pos12_rev | This work | Mutagenic PCR primer | ATCTACCGCTAACGCCGC |
| Sequence-based reagent | WT_DHFR_pos13_rev | This work | Mutagenic PCR primer | GCGATCTACCGCTAACGC |
| Sequence-based reagent | WT_DHFR_pos14_rev | This work | Mutagenic PCR primer | AACGCGATCTACCGCTAAC |
| Sequence-based reagent | WT_DHFR_pos15_rev | This work | Mutagenic PCR primer | GATAACGCGATCTACCGCTAAC |
| Sequence-based reagent | WT_DHFR_pos16_rev | This work | Mutagenic PCR primer | GCCGATAACGCGATCTACC |
| Sequence-based reagent | WT_DHFR_pos17_rev | This work | Mutagenic PCR primer | CATGCCGATAACGCGATCTAC |
| Sequence-based reagent | WT_DHFR_pos18_rev | This work | Mutagenic PCR primer | TTCCATGCCGATAACGCG |
| Sequence-based reagent | WT_DHFR_pos19_rev | This work | Mutagenic PCR primer | GTTTTCCATGCCGATAACGC |
| Sequence-based reagent | WT_DHFR_pos20_rev | This work | Mutagenic PCR primer | GGCGTTTTCCATGCCGATAACG |
| Sequence-based reagent | WT_DHFR_pos21_rev | This work | Mutagenic PCR primer | CATGGCGTTTTCCATGCC |
| Sequence-based reagent | WT_DHFR_pos22_rev | This work | Mutagenic PCR primer | CGGCATGGCGTTTTCCAT |
| Sequence-based reagent | WT_DHFR_pos22_rev2 | This work | Mutagenic PCR primer | CGGCATGGCGTTTTCCATG |
| Sequence-based reagent | WT_DHFR_pos23_rev | This work | Mutagenic PCR primer | CCACGGCATGGCGTTTTC |
| Sequence-based reagent | WT_DHFR_pos24_rev | This work | Mutagenic PCR primer | GTTCCACGGCATGGCGTT |
| Sequence-based reagent | WT_DHFR_pos25_rev | This work | Mutagenic PCR primer | CAGGTTCCACGGCATGGC |
| Sequence-based reagent | WT_DHFR_pos26_rev | This work | Mutagenic PCR primer | AGGCAGGTTCCACGGCAT |
| Sequence-based reagent | WT_DHFR_pos27_rev | This work | Mutagenic PCR primer | GGCAGGCAGGTTCCACGG |
| Sequence-based reagent | WT_DHFR_pos28_rev | This work | Mutagenic PCR primer | ATCGGCAGGCAGGTTCCA |
| Sequence-based reagent | WT_DHFR_pos29_rev | This work | Mutagenic PCR primer | GAGATCGGCAGGCAGGTT |
| Sequence-based reagent | WT_DHFR_pos30_rev | This work | Mutagenic PCR primer | GGCGAGATCGGCAGGCAG |
| Sequence-based reagent | WT_DHFR_pos31_rev | This work | Mutagenic PCR primer | CCAGGCGAGATCGGCAGG |
| Sequence-based reagent | WT_DHFR_pos32_rev | This work | Mutagenic PCR primer | AAACCAGGCGAGATCGGC |
| Sequence-based reagent | WT_DHFR_pos33_rev | This work | Mutagenic PCR primer | TTTAAACCAGGCGAGATCGG |
| Sequence-based reagent | WT_DHFR_pos34_rev | This work | Mutagenic PCR primer | GCGTTTAAACCAGGCGAGAT |
| Sequence-based reagent | WT_DHFR_pos35_rev | This work | Mutagenic PCR primer | GTTGCGTTTAAACCAGGCGA |
| Sequence-based reagent | WT_DHFR_pos36_rev | This work | Mutagenic PCR primer | GGTGTTGCGTTTAAACCAGG |
| Sequence-based reagent | WT_DHFR_pos37_rev | This work | Mutagenic PCR primer | TAAGGTGTTGCGTTTAAACCAGG |
| Sequence-based reagent | WT_DHFR_pos38_rev | This work | Mutagenic PCR primer | ATTTAAGGTGTTGCGTTTAAACCAGG |
| Sequence-based reagent | WT_DHFR_pos39_rev | This work | Mutagenic PCR primer | TTTATTTAAGGTGTTGCGTTTAAACCAG |
| Sequence-based reagent | WT_DHFR_pos40_rev | This work | Mutagenic PCR primer | GGGTTTATTTAAGGTGTTGCGTTTAAAC |
| Sequence-based reagent | WT_DHFR_pos41_rev | This work | Mutagenic PCR primer | CACGGGTTTATTTAAGGTGTTGCGT |
| Sequence-based reagent | WT_DHFR_pos42_rev | This work | Mutagenic PCR primer | AATCACGGGTTTATTTAAGGTGTTGC |
| Sequence-based reagent | WT_DHFR_pos42_rev2 | This work | Mutagenic PCR primer | AATCACGGGTTTATTTAAGGTGTTGC |
| Sequence-based reagent | WT_DHFR_pos43_rev | This work | Mutagenic PCR primer | CATAATCACGGGTTTATTTAAGGTGTTG |
| Sequence-based reagent | WT_DHFR_pos43_rev2 | This work | Mutagenic PCR primer | CATAATCACGGGTTTATTTAAGGTGTTG |
| Sequence-based reagent | WT_DHFR_pos44_rev | This work | Mutagenic PCR primer | GCCCATAATCACGGGTTTATTTAAGG |
| Sequence-based reagent | WT_DHFR_pos45_rev | This work | Mutagenic PCR primer | GCGGCCCATAATCACGGG |
| Sequence-based reagent | WT_DHFR_pos46_rev | This work | Mutagenic PCR primer | ATGGCGGCCCATAATCAC |
| Sequence-based reagent | WT_DHFR_pos47_rev | This work | Mutagenic PCR primer | GGTATGGCGGCCCATAATC |
| Sequence-based reagent | WT_DHFR_pos48_rev | This work | Mutagenic PCR primer | CCAGGTATGGCGGCCCATA |
| Sequence-based reagent | WT_DHFR_pos49_rev | This work | Mutagenic PCR primer | TTCCCAGGTATGGCGGCC |
| Sequence-based reagent | WT_DHFR_pos50_rev | This work | Mutagenic PCR primer | TGATTCCCAGGTATGGCGGC |
| Sequence-based reagent | WT_DHFR_pos51_rev | This work | Mutagenic PCR primer | GATTGATTCCCAGGTATGGCGG |
| Sequence-based reagent | WT_DHFR_pos52_rev | This work | Mutagenic PCR primer | ACCGATTGATTCCCAGGTATG |
| Sequence-based reagent | WT_DHFR_pos53_rev | This work | Mutagenic PCR primer | ACGACCGATTGATTCCCAG |
| Sequence-based reagent | WT_DHFR_pos54_rev | This work | Mutagenic PCR primer | CGGACGACCGATTGATTCC |
| Sequence-based reagent | WT_DHFR_pos55_rev | This work | Mutagenic PCR primer | CAACGGACGACCGATTGATTC |
| Sequence-based reagent | WT_DHFR_pos56_rev | This work | Mutagenic PCR primer | TGGCAACGGACGACCGAT |
| Sequence-based reagent | WT_DHFR_pos57_rev | This work | Mutagenic PCR primer | TCCTGGCAACGGACGACC |
| Sequence-based reagent | WT_DHFR_pos58_rev | This work | Mutagenic PCR primer | GCGTCCTGGCAACGGACG |
| Sequence-based reagent | WT_DHFR_pos59_rev | This work | Mutagenic PCR primer | TTTGCGTCCTGGCAACGG |
| Sequence-based reagent | WT_DHFR_pos60_rev | This work | Mutagenic PCR primer | ATTTTTGCGTCCTGGCAAC |
| Sequence-based reagent | WT_DHFR_pos61_rev | This work | Mutagenic PCR primer | AATATTTTTGCGTCCTGGCAAC |
| Sequence-based reagent | WT_DHFR_pos62_rev | This work | Mutagenic PCR primer | GATAATATTTTTGCGTCCTGGCAAC |
| Sequence-based reagent | WT_DHFR_pos63_rev | This work | Mutagenic PCR primer | GAGGATAATATTTTTGCGTCCTGGC |
| Sequence-based reagent | WT_DHFR_pos64_rev | This work | Mutagenic PCR primer | GCTGAGGATAATATTTTTGCGTCCTG |
| Sequence-based reagent | WT_DHFR_pos65_rev | This work | Mutagenic PCR primer | ACTGCTGAGGATAATATTTTTGCGTCCT |
| Sequence-based reagent | WT_DHFR_pos66_rev | This work | Mutagenic PCR primer | TTGACTGCTGAGGATAATATTTTTGCG |
| Sequence-based reagent | WT_DHFR_pos66_rev2 | This work | Mutagenic PCR primer | TTGACTGCTGAGGATAATATTTTTGC |
| Sequence-based reagent | WT_DHFR_pos67_rev | This work | Mutagenic PCR primer | CGGTTGACTGCTGAGGATAATATTTTTG |
| Sequence-based reagent | WT_DHFR_pos67_rev2 | This work | Mutagenic PCR primer | CGGTTGACTGCTGAGGATAATATTTTTG |
| Sequence-based reagent | WT_DHFR_pos68_rev | This work | Mutagenic PCR primer | ACCCGGTTGACTGCTGAG |
| Sequence-based reagent | WT_DHFR_pos69_rev | This work | Mutagenic PCR primer | CGTACCCGGTTGACTGCT |
| Sequence-based reagent | WT_DHFR_pos70_rev | This work | Mutagenic PCR primer | GTCCGTACCCGGTTGACT |
| Sequence-based reagent | WT_DHFR_pos71_rev | This work | Mutagenic PCR primer | ATCGTCCGTACCCGGTTG |
| Sequence-based reagent | WT_DHFR_pos72_rev | This work | Mutagenic PCR primer | GCGATCGTCCGTACCCGG |
| Sequence-based reagent | WT_DHFR_pos73_rev | This work | Mutagenic PCR primer | TACGCGATCGTCCGTACC |
| Sequence-based reagent | WT_DHFR_pos73_rev2 | This work | Mutagenic PCR primer | TACGCGATCGTCCGTACC |
| Sequence-based reagent | WT_DHFR_pos74_rev | This work | Mutagenic PCR primer | CGTTACGCGATCGTCCGT |
| Sequence-based reagent | WT_DHFR_pos74_rev2 | This work | Mutagenic PCR primer | CGTTACGCGATCGTCCGTAC |
| Sequence-based reagent | WT_DHFR_pos75_rev | This work | Mutagenic PCR primer | CCACGTTACGCGATCGTC |
| Sequence-based reagent | WT_DHFR_pos76_rev | This work | Mutagenic PCR primer | CACCCACGTTACGCGATC |
| Sequence-based reagent | WT_DHFR_pos77_rev | This work | Mutagenic PCR primer | CTTCACCCACGTTACGCG |
| Sequence-based reagent | WT_DHFR_pos78_rev | This work | Mutagenic PCR primer | CGACTTCACCCACGTTACG |
| Sequence-based reagent | WT_DHFR_pos79_rev | This work | Mutagenic PCR primer | CACCGACTTCACCCACGTTAC |
| Sequence-based reagent | WT_DHFR_pos80_rev | This work | Mutagenic PCR primer | ATCCACCGACTTCACCCACGTTAC |
| Sequence-based reagent | WT_DHFR_pos80_rev2 | This work | Mutagenic PCR primer | ATCCACCGACTTCACCCAC |
| Sequence-based reagent | WT_DHFR_pos81_rev | This work | Mutagenic PCR primer | TTCATCCACCGACTTCACCCA |
| Sequence-based reagent | WT_DHFR_pos82_rev | This work | Mutagenic PCR primer | GGCTTCATCCACCGACTTCAC |
| Sequence-based reagent | WT_DHFR_pos82_rev2 | This work | Mutagenic PCR primer | GGCTTCATCCACCGACTTCAC |
| Sequence-based reagent | WT_DHFR_pos83_rev | This work | Mutagenic PCR primer | GATGGCTTCATCCACCGAC |
| Sequence-based reagent | WT_DHFR_pos84_rev | This work | Mutagenic PCR primer | CGCGATGGCTTCATCCAC |
| Sequence-based reagent | WT_DHFR_pos84_rev2 | This work | Mutagenic PCR primer | CGCGATGGCTTCATCCAC |
| Sequence-based reagent | WT_DHFR_pos85_rev | This work | Mutagenic PCR primer | CGCCGCGATGGCTTCATC |
| Sequence-based reagent | WT_DHFR_pos86_rev | This work | Mutagenic PCR primer | ACACGCCGCGATGGCTTC |
| Sequence-based reagent | WT_DHFR_pos87_rev | This work | Mutagenic PCR primer | ACCACACGCCGCGATGGC |
| Sequence-based reagent | WT_DHFR_pos88_rev | This work | Mutagenic PCR primer | GTCACCACACGCCGCGAT |
| Sequence-based reagent | WT_DHFR_pos89_rev | This work | Mutagenic PCR primer | TACGTCACCACACGCCGC |
| Sequence-based reagent | WT_DHFR_pos89_rev2 | This work | Mutagenic PCR primer | TACGTCACCACACGCCG |
| Sequence-based reagent | WT_DHFR_pos90_rev | This work | Mutagenic PCR primer | TGGTACGTCACCACACGC |
| Sequence-based reagent | WT_DHFR_pos91_rev | This work | Mutagenic PCR primer | TTCTGGTACGTCACCACACGC |
| Sequence-based reagent | WT_DHFR_pos92_rev | This work | Mutagenic PCR primer | GATTTCTGGTACGTCACCACACG |
| Sequence-based reagent | WT_DHFR_pos93_rev | This work | Mutagenic PCR primer | CATGATTTCTGGTACGTCACCACAC |
| Sequence-based reagent | WT_DHFR_pos94_rev | This work | Mutagenic PCR primer | CACCATGATTTCTGGTACGTCACC |
| Sequence-based reagent | WT_DHFR_pos95_rev | This work | Mutagenic PCR primer | AATCACCATGATTTCTGGTACGTCA |
| Sequence-based reagent | WT_DHFR_pos95_rev2 | This work | Mutagenic PCR primer | AATCACCATGATTTCTGGTACGTC |
| Sequence-based reagent | WT_DHFR_pos96_rev | This work | Mutagenic PCR primer | GCCAATCACCATGATTTCTGGTAC |
| Sequence-based reagent | WT_DHFR_pos97_rev | This work | Mutagenic PCR primer | GCCGCCAATCACCATGATTT |
| Sequence-based reagent | WT_DHFR_pos98_rev | This work | Mutagenic PCR primer | ACCGCCGCCAATCACCATGATTTC |
| Sequence-based reagent | WT_DHFR_pos99_rev | This work | Mutagenic PCR primer | GCGACCGCCGCCAATCAC |
| Sequence-based reagent | WT_DHFR_pos100_rev | This work | Mutagenic PCR primer | AACGCGACCGCCGCCAAT |
| Sequence-based reagent | WT_DHFR_pos101_rev | This work | Mutagenic PCR primer | ATAAACGCGACCGCCGCC |
| Sequence-based reagent | WT_DHFR_pos102_rev | This work | Mutagenic PCR primer | TTCATAAACGCGACCGCC |
| Sequence-based reagent | WT_DHFR_pos103_rev | This work | Mutagenic PCR primer | CTGTTCATAAACGCGACCG |
| Sequence-based reagent | WT_DHFR_pos104_rev | This work | Mutagenic PCR primer | GAACTGTTCATAAACGCGACC |
| Sequence-based reagent | WT_DHFR_pos104_rev2 | This work | Mutagenic PCR primer | GAACTGTTCATAAACGCGACCG |
| Sequence-based reagent | WT_DHFR_pos105_rev | This work | Mutagenic PCR primer | CAAGAACTGTTCATAAACGCGAC |
| Sequence-based reagent | WT_DHFR_pos106_rev | This work | Mutagenic PCR primer | TGGCAAGAACTGTTCATAAACGC |
| Sequence-based reagent | WT_DHFR_pos107_rev | This work | Mutagenic PCR primer | TTTTGGCAAGAACTGTTCATAAACG |
| Sequence-based reagent | WT_DHFR_pos107_rev2 | This work | Mutagenic PCR primer | TTTTGGCAAGAACTGTTCATAAACG |
| Sequence-based reagent | WT_DHFR_pos108_rev | This work | Mutagenic PCR primer | CGCTTTTGGCAAGAACTGTTCATAAA |
| Sequence-based reagent | WT_DHFR_pos109_rev | This work | Mutagenic PCR primer | TTGCGCTTTTGGCAAGAACT |
| Sequence-based reagent | WT_DHFR_pos110_rev | This work | Mutagenic PCR primer | TTTTTGCGCTTTTGGCAAGAAC |
| Sequence-based reagent | WT_DHFR_pos111_rev | This work | Mutagenic PCR primer | CAGTTTTTGCGCTTTTGGCAAG |
| Sequence-based reagent | WT_DHFR_pos112_rev | This work | Mutagenic PCR primer | ATACAGTTTTTGCGCTTTTGGCAA |
| Sequence-based reagent | WT_DHFR_pos113_rev | This work | Mutagenic PCR primer | CAGATACAGTTTTTGCGCTTTTGG |
| Sequence-based reagent | WT_DHFR_pos114_rev | This work | Mutagenic PCR primer | CGTCAGATACAGTTTTTGCGCTTTT |
| Sequence-based reagent | WT_DHFR_pos115_rev | This work | Mutagenic PCR primer | ATGCGTCAGATACAGTTTTTGCG |
| Sequence-based reagent | WT_DHFR_pos116_rev | This work | Mutagenic PCR primer | GATATGCGTCAGATACAGTTTTTGCG |
| Sequence-based reagent | WT_DHFR_pos117_rev | This work | Mutagenic PCR primer | GTCGATATGCGTCAGATACAGTTTTTG |
| Sequence-based reagent | WT_DHFR_pos118_rev | This work | Mutagenic PCR primer | TGCGTCGATATGCGTCAGATA |
| Sequence-based reagent | WT_DHFR_pos118_rev2 | This work | Mutagenic PCR primer | TGCGTCGATATGCGTCAGATAC |
| Sequence-based reagent | WT_DHFR_pos119_rev | This work | Mutagenic PCR primer | TTCTGCGTCGATATGCGTCA |
| Sequence-based reagent | WT_DHFR_pos120_rev | This work | Mutagenic PCR primer | CACTTCTGCGTCGATATGCG |
| Sequence-based reagent | WT_DHFR_pos121_rev | This work | Mutagenic PCR primer | TTCCACTTCTGCGTCGATATG |
| Sequence-based reagent | WT_DHFR_pos122_rev | This work | Mutagenic PCR primer | GCCTTCCACTTCTGCGTC |
| Sequence-based reagent | WT_DHFR_pos123_rev | This work | Mutagenic PCR primer | GTCGCCTTCCACTTCTGC |
| Sequence-based reagent | WT_DHFR_pos124_rev | This work | Mutagenic PCR primer | GGTGTCGCCTTCCACTTC |
| Sequence-based reagent | WT_DHFR_pos125_rev | This work | Mutagenic PCR primer | ATGGGTGTCGCCTTCCAC |
| Sequence-based reagent | WT_DHFR_pos126_rev | This work | Mutagenic PCR primer | GAAATGGGTGTCGCCTTCC |
| Sequence-based reagent | WT_DHFR_pos127_rev | This work | Mutagenic PCR primer | CGGGAAATGGGTGTCGCC |
| Sequence-based reagent | WT_DHFR_pos128_rev | This work | Mutagenic PCR primer | ATCCGGGAAATGGGTGTC |
| Sequence-based reagent | WT_DHFR_pos129_rev | This work | Mutagenic PCR primer | GTAATCCGGGAAATGGGTGTC |
| Sequence-based reagent | WT_DHFR_pos130_rev | This work | Mutagenic PCR primer | CTCGTAATCCGGGAAATGGG |
| Sequence-based reagent | WT_DHFR_pos131_rev | This work | Mutagenic PCR primer | CGGCTCGTAATCCGGGAA |
| Sequence-based reagent | WT_DHFR_pos131_rev2 | This work | Mutagenic PCR primer | CGGCTCGTAATCCGGGAAATG |
| Sequence-based reagent | WT_DHFR_pos132_rev | This work | Mutagenic PCR primer | ATCCGGCTCGTAATCCGG |
| Sequence-based reagent | WT_DHFR_pos133_rev | This work | Mutagenic PCR primer | GTCATCCGGCTCGTAATCC |
| Sequence-based reagent | WT_DHFR_pos134_rev | This work | Mutagenic PCR primer | CCAGTCATCCGGCTCGTA |
| Sequence-based reagent | WT_DHFR_pos135_rev | This work | Mutagenic PCR primer | TTCCCAGTCATCCGGCTC |
| Sequence-based reagent | WT_DHFR_pos135_rev2 | This work | Mutagenic PCR primer | TTCCCAGTCATCCGGCTC |
| Sequence-based reagent | WT_DHFR_pos136_rev | This work | Mutagenic PCR primer | CGATTCCCAGTCATCCGG |
| Sequence-based reagent | WT_DHFR_pos136_rev2 | This work | Mutagenic PCR primer | CGATTCCCAGTCATCCGGC |
| Sequence-based reagent | WT_DHFR_pos137_rev | This work | Mutagenic PCR primer | TACCGATTCCCAGTCATCCG |
| Sequence-based reagent | WT_DHFR_pos138_rev | This work | Mutagenic PCR primer | GAATACCGATTCCCAGTCATCC |
| Sequence-based reagent | WT_DHFR_pos139_rev | This work | Mutagenic PCR primer | GCTGAATACCGATTCCCAGTC |
| Sequence-based reagent | WT_DHFR_pos140_rev | This work | Mutagenic PCR primer | TTCGCTGAATACCGATTCCCA |
| Sequence-based reagent | WT_DHFR_pos140_rev2 | This work | Mutagenic PCR primer | TTCGCTGAATACCGATTCCCAG |
| Sequence-based reagent | WT_DHFR_pos141_rev | This work | Mutagenic PCR primer | GAATTCGCTGAATACCGATTCCC |
| Sequence-based reagent | WT_DHFR_pos142_rev | This work | Mutagenic PCR primer | GTGGAATTCGCTGAATACCGATTC |
| Sequence-based reagent | WT_DHFR_pos143_rev | This work | Mutagenic PCR primer | ATCGTGGAATTCGCTGAATACC |
| Sequence-based reagent | WT_DHFR_pos144_rev | This work | Mutagenic PCR primer | AGCATCGTGGAATTCGCTG |
| Sequence-based reagent | WT_DHFR_pos145_rev | This work | Mutagenic PCR primer | ATCAGCATCGTGGAATTCGC |
| Sequence-based reagent | WT_DHFR_pos146_rev | This work | Mutagenic PCR primer | CGCATCAGCATCGTGGAATT |
| Sequence-based reagent | WT_DHFR_pos147_rev | This work | Mutagenic PCR primer | CTGCGCATCAGCATCGTG |
| Sequence-based reagent | WT_DHFR_pos148_rev | This work | Mutagenic PCR primer | GTTCTGCGCATCAGCATC |
| Sequence-based reagent | WT_DHFR_pos149_rev | This work | Mutagenic PCR primer | AGAGTTCTGCGCATCAGC |
| Sequence-based reagent | WT_DHFR_pos150_rev | This work | Mutagenic PCR primer | GTGAGAGTTCTGCGCATCAG |
| Sequence-based reagent | WT_DHFR_pos151_rev | This work | Mutagenic PCR primer | GCTGTGAGAGTTCTGCGC |
| Sequence-based reagent | WT_DHFR_pos152_rev | This work | Mutagenic PCR primer | ATAGCTGTGAGAGTTCTGCG |
| Sequence-based reagent | WT_DHFR_pos153_rev | This work | Mutagenic PCR primer | GCAATAGCTGTGAGAGTTCTGC |
| Sequence-based reagent | WT_DHFR_pos154_rev | This work | Mutagenic PCR primer | AAAGCAATAGCTGTGAGAGTTCTG |
| Sequence-based reagent | WT_DHFR_pos155_rev | This work | Mutagenic PCR primer | CTCAAAGCAATAGCTGTGAGAGTTC |
| Sequence-based reagent | WT_DHFR_pos156_rev | This work | Mutagenic PCR primer | AATCTCAAAGCAATAGCTGTGAGAGTTC |
| Sequence-based reagent | WT_DHFR_pos157_rev | This work | Mutagenic PCR primer | CAGAATCTCAAAGCAATAGCTGTGAGAG |
| Sequence-based reagent | WT_DHFR_pos158_rev | This work | Mutagenic PCR primer | CTCCAGAATCTCAAAGCAATAGCTG |
| Sequence-based reagent | WT_DHFR_pos159_rev | This work | Mutagenic PCR primer | CCGCTCCAGAATCTCAAAGC |
| Sequence-based reagent | SL1_FWD | This work | Round one amplicon PCR primer | CACTCTTTCCCTACACGACGCTCTTCCGATCTNNNNACTTTAATAACGAGATATACCATG |
| Sequence-based reagent | SL1_REV | This work | Round one amplicon PCR primer | TGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNGTATGGCGGCCCATAAT |
| Sequence-based reagent | SL2_FWD | This work | Round one amplicon PCR primer | CACTCTTTCCCTACACGACGCTCTTCCGATCTNNNNACACCTTAAATAAACCCGTG |
| Sequence-based reagent | SL2_REV | This work | Round one amplicon PCR primer |
TGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNCACGCCGCGATGGC |
| Sequence-based reagent | SL3_FWD | This work | Round one amplicon PCR primer | CACTCTTTCCCTACACGACGCTCTTCCGATCTNNNNTGAAGTCGGTGGATGAA |
| Sequence-based reagent | SL3_REV | This work | Round one amplicon PCR primer | TGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNGAAATGGGTGTCGCC |
| Sequence-based reagent | SL4_FWD | This work | Round one amplicon PCR primer | CACTCTTTCCCTACACGACGCTCTTCCGATCTNNNNCGACGCAGAAGTGGAA |
| Sequence-based reagent | SL4_REV | This work | Round one amplicon PCR primer | TGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNGCTTGTCGACGCCTG |
| Sequence-based reagent | D501 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | AATGATACGGCGACCACCGAGATCTACACTATAGCCTACACTCTTTCCCTACACGAC |
| Sequence-based reagent | D502 | Illumina/Reynolds et al. Cell 2011 | Round two amplicon PCR primer | AATGATACGGCGACCACCGAGATCTACACATAGAGGCACACTCTTTCCCTACACGAC |
| Sequence-based reagent | D503 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer |
AATGATACGGCGACCACCGAGATCTACACCCTATCCTACACTCTTT
CCCTACACGAC |
| Sequence-based reagent | D504 | Illumina/Reynolds et al. Cell 2011 | Round two amplicon PCR primer | AATGATACGGCGACCACCGAGATCTACACGGCTCTGAACACTCTTTCCCTACACGAC |
| Sequence-based reagent | D505 | Illumina/Reynolds et al. Cell 2011 | Round two amplicon PCR primer | AATGATACGGCGACCACCGAGATCTACACAGGCGAAGACACTCTTTCCCTACACGAC |
| Sequence-based reagent | D506 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | AATGATACGGCGACCACCGAGATCTACACTAATCTTAACACTCTTTCCCTACACGAC |
| Sequence-based reagent | D507 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | AATGATACGGCGACCACCGAGATCTACACCAGGACGTACACTCTTTCCCTACACGAC |
| Sequence-based reagent | D508 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | AATGATACGGCGACCACCGAGATCTACACGTACTGACACACTCTTTCCCTACACGAC |
| Sequence-based reagent | D701 | Illumina/Reynolds et al. Cell 2011 | Round two amplicon PCR primer | CAAGCAGAAGACGGCATACGAGATCGAGTAATGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | D702 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | CAAGCAGAAGACGGCATACGAGATTCTCCGGAGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | D703 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | CAAGCAGAAGACGGCATACGAGATAATGAGCGGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | D704 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | CAAGCAGAAGACGGCATACGAGATGGAATCTCGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | D705 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | CAAGCAGAAGACGGCATACGAGATTTCTGAATGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | D706 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | CAAGCAGAAGACGGCATACGAGATACGAATTCGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | D707 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | CAAGCAGAAGACGGCATACGAGATAGCTTCAGGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | D708 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer |
CAAGCAGAAGACGGCATACGAG
ATGCGCATTAGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | D709 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | CAAGCAGAAGACGGCATACGAGATCATAGCCGGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | D710 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | CAAGCAGAAGACGGCATACGAGATTTCGCGGAGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | D711 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | CAAGCAGAAGACGGCATACGAGATGCGCGAGAGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | D712 | Illumina/Reynolds et al., 2011 | Round two amplicon PCR primer | CAAGCAGAAGACGGCATACGAGATCTATCGCTGTGACTGGAGTTCAGACGTG |
| Sequence-based reagent | KanSacB_round1_fwd | This work | PCR primer | caggcatctggtgaataaTCCTTTTATGATTTTCTATCAAACAAAAGAGG |
| Sequence-based reagent | KanSacB_round1_rev | This work | PCR primer | tcaatgcgttcagaacgctcaggattcatGCTTGGTCGGTCATTTCGAAC |
| Sequence-based reagent | KanSacB_round2_fwd/Anderson_promoter_outer_fwd | This work | PCR primer | gtcaaagcaaaccgttgctgatttatggcaagccggaagcgcaacaggcatctggtgaataa |
| Sequence-based reagent | KanSacB_round2_rev/Anderson_promoter_outer_rev | This work | PCR primer | ccaccacatcgcgcagcggcaatacggggatttcaatgcgttcagaacgctcaggattcat |
| Sequence-based reagent | Anderson_promoter_outer_fwd/KanSacB_round2_fwd | This work | PCR primer | same as KanSacB_round2_fwd/Anderson_promoter_outer_fwd |
| Sequence-based reagent | Anderson_promoter_inner_fwd | This work | PCR primer | CCTAGGACTGAGCTAGCTGTCAAcgtcagtatatggggatgtttcccc |
| Sequence-based reagent | Anderson_promoter_inner_rev | This work | PCR primer | GCTAGCTCAGTCCTAGGTATAATGCTAGCAGGAtacctggcggaaattaaactaagagag |
| Sequence-based reagent | Anderson_promoter_outer_rev/KanSacB_round2_rev | This work | PCR primer | same as KanSacB_round2_rev/Anderson_promoter_outer_rev |