TABLE 3.
Primers and crRNAs used in this study
| Primer | Sequence (5′–3′)a |
|---|---|
| pyrG Fw 2g08660 | TTCCCCCTAATCACTGCACCCTTTCCCCGGACTCTGCACGgaattctcatgtttgacagc |
| pyrG Rv 2g08660 | GTGATATAGAGACGAGGACAAATATGCTAGAAGCTTTATGggatccacaggacgggtgtgg |
| 2g08660 Fw Scr | GACTCAGACCCGCTTCGC |
| 2g08660 Rv Scr | CCACGCACGATATCCATGACC |
| pyrG Fw 2g08670 | TGCGTGGGAATTGTATAATTTATAATAAAACTGCAAGCATgaattctcatgtttgacagc |
| pyrG Rv 2g08670 | ACGGGGCGGTTCTATGTTTCATTCTATTTTCATGGTGAGCggatccacaggacgggtgtgg |
| 2g08670 Fw Scr | ACTTTCGCGTCCTCACCTC |
| 2g08670 Rv Scr | CCTAACGCACCCGCCTTG |
| pyrG Fw 3g13230 | CGGCACGATCGCCCCTCTTCATGAACCGCATTGTTTGTTGgaattctcatgtttgacagc |
| pyrG Rv 3g13230 | GATCTCAAAGTCGAGGGTGATGGATGAACGGTCATCGACGggatccacaggacgggtgtgg |
| 3g13230 Fw Scr | GGTGCGTTGGCAATACAACC |
| 3g13230 Rv Scr | ACCGGATCCGTAATAGTCCG |
| pyrG Fw 3g13260 | TCTGTTTTTTAGGCTTTCACTTGTGGTCTCTGGTCTGTCTgaattctcatgtttgacagc |
| pyrG Rv 3g13260 | TAACAAATCAACTTTGAAGTCGGCAAACATCAACTCAAGTggatccacaggacgggtgtgg |
| 3g13260 Fw Scr | GCCTGCTTCACAGTAGTCGAG |
| 3g13260 Rv Scr | CATAGACGCCAGAACACCGC |
| pyrG Fw 3g13270 | ACCTCTGTCCGGGCTGATACCGGCTCCGGGTCTTCCGCCTgaattctcatgtttgacagc |
| pyrG Rv 3g13270 | AGCGCGTCAGAGAGAAATTGACAGGCCCGACCTCGGCATTggatccacaggacgggtgtgg |
| 3g13270 Fw Scr | TGCGTCGTTTGCAGCTGG |
| 3g13270 Rv Scr | CCATTGCGGATGTCCCATCG |
| pyrG Fw 3g13300 | GCCTTGGTGCCGTTCTTTCAGTCGGTTGTTGTTCTGCTTTgaattctcatgtttgacagc |
| pyrG Rv 3g13300 | TATCCAGAGCCTTTCGTCACCCTGATGACATGACTTGAGggatccacaggacgggtgtgg |
| 3g13300 Fw Scr | CGTCTCAAATCACGTCGCGG |
| 3g13300 Rv Scr | ACAGGCTATGCTCAGTAATCGG |
| pyrG Fw 4g07080 | TAATTTTGTCCTTCGCTGATCGGCTATAACGTGGCATCAGgaattctcatgtttgacagc |
| pyrG Rv 4g07080 | AGAGTTCATCCCAGACGATGGGTAGATTAGATTCTGAGGAggatccacaggacgggtgtgg |
| 4g07080 Fw Scr | ACCTCCCCTTAACTCGTCACC |
| 4g07080 Rv Scr | CGGTCTGCTAACGTAAGCCG |
| pyrG Fw 7g01440 | GCAGCACGCATCTCAGCATTAGTACTCAACAGATGGAAACgaattctcatgtttgacagc |
| pyrG Rv 7g01440 | TATAGCCTTGGGAAAGGGTAGATGGTAGAGGATACCGTTTggatccacaggacgggtgtgg |
| 7g01440 Fw Scr | TTCAGCAGTGTCAGCACTCGG |
| 7g01440 Rv Scr | AGCCATCGACGTTCACGC |
| pyrG Fw 7g01560 | TCTATAAGTTGTTCTAAGCCCATAACGCACCATCATGACCgaattctcatgtttgacagc |
| pyrG Rv 7g01560 | TGGTCCAAGATCAATATAAGACAGCATATCGTTAATGAAGggatccacaggacgggtgtgg |
| 7g01560 Fw Scr | GCAACGTCTGCCGCGATG |
| 7g01560 Rv Scr | TCAGTTGGTGGCACAGTCC |
| pyrG Fw 8g06360 | TTCATGGTATGATCATGCTGACTTTGCTCGAACTTAATGAgaattctcatgtttgacagc |
| pyrG Rv 8g06360 | AAGAACGGCTGCTCTTGTATGGTGTGCATCTACAATGCTTggatccacaggacgggtgtgg |
| 8g06360 Fw Scr | GCCTGGCAGATGCAAAGGC |
| 8g06360 Rv Scr | AGTACCCTCGGCAAGCGC |
| Ap pyrG Fw scr | gcccttgcagagaagcac |
| Ap pyrG Rv scr | cagcataaattccacgaccagc |
| Hyg Fw 3g13230 | GCGCGACGCGATTCGCGGCACGATCGCCCCTCTTCATGAACCGagcttgcatgcctgcaggtcg |
| hspA Rv 3g13230 | CCAAGAATGTCTGGAGAAGAGCTGGAACTTTGAGAGAACATtgtgaagaagtgaggaggg |
| Hyg Fw 4g07080 | GAAACTATACTCATAATTTTGTCCTTCGCTGATCGGCTATAAagcttgcatgcctgcaggtcg |
| hspA Rv 4g07080 | CTCCGATGTAGATATCGTGTCCTGGGATCTTGTTCATAGCCATtgtgaagaagtgaggaggg |
| 2g08660 5 gRNA | AGATGTCCGTTCTCCGTCAT |
| 2g08660 3 gRNA | TCGCAGCCTAGAAAGACAGC |
| 2g08670 5 gRNA | TAATAAAACTGCAAGCATCA |
| 2g08670 3 gRNA | GGAGGCCGTGCTCAAGTACC |
| 3g13230 5 gRNA | GAACCGCATTGTTTGTTGCA |
| 3g13230 3 gRNA | GAGGAGCCGCTCTCACGGCG |
| 3g13260 5 gRNA | GTGGTCTCTGGTCTGTCTCA |
| 3g13260 3 gRNA | CATCCCCCGTGTAAGCATTT |
| 3g13270 5 gRNA | GCTCCGGGTCTTCCGCCTTT |
| 3g13270 3 gRNA | TAGACATATACCTCTTTCCT |
| 3g13300 5 gRNA | GTCTACAGTCAAGATGCGTA |
| 3g13300 3 gRNA | ATTTACGAAAGCTCAAGCCT |
| 4g07080 5 gRNA | GCTATAACGTGGCATCAGCA |
| 4g07080 3 gRNA | GGCCGCGGCAATCTTTGACG |
| 7g01440 5 gRNA | TACTCAACAGATGGAAACAT |
| 7g01440 3 gRNA | AAGAATGGTTAATCATCATT |
| 7g01560 5 gRNA | TAACGCACCATCATGACCCG |
| 7g01560 3 gRNA | AGAGAACTAGCGGAGGATTC |
| 8g06360 5 gRNA | AAGTGCTAAGAATGCATTTA |
| 8g06360 3 gRNA | ACAAGCATTGTAGATGCACA |
a
Sequences for the amplification of the A. parasiticus pyrG cassette are indicated in lowercase, while the microhomology arms for each gene are indicated in uppercase.