TABLE 1 .
Oligonucleotides and crRNAs used for this study
| Type, purpose, and name | Sequencea |
|---|---|
| Oligonucleotides for | |
| Cloning HygR cassette into pCR-Blunt II-TOPO |
|
| gpdA(p)-For | CGAGCTCCCAAATCTGTCC |
| trpC(t)-Rev | AGCTTGCATGCCTGCAGG |
| Amplification of HygR cassette with 35-bp microhomology |
|
| 35 bp-pksP-HygR-F | ACTCCTTGCAGTCACGGCCGCGCAAACCACTCGCCCGAGCTCCCAAATCTGTCCA |
| 35 bp-pksP-HygR-R | AGAAACCAATTTAGGTATGCACAACTCCACCCCCAAGCTTGCATGCCTGCAGGTC |
|
Amplification of HygR cassette with 50-bp microhomology |
|
| 50 bp-pksP-HygR-F | TCCAGGATAATCACTACTCCTTGCAGTCACGGCCGCGCAAACCACTCGCCCGAGCTCCCAAATCTGTCCA |
| 50 bp-pksP-HygR-R | CAAGGAGAAAACAAAAGAAACCAATTTAGGTATGCACAACTCCACCCCCAAGCTTGCATGCCTGCAGGTC |
| Amplification of Southern blot probe | |
| pksP-Probe-Forward | GGCACGCTGTGCAGGACG |
| pksP-Probe-Reverse | GCCCGTCATCCCACTTGC |
| Sequencing DI15-102 isolate | |
| pksP(p)-159-for | TGAAGAGGTTGGACCCCAGTCGT |
| pksP-141-Rev | TTCCTGACGCAGTGCATGGAAGC |
| pksP-6521-For | CCAAACAAATGGGATACTTTGGTGGGCC |
| pksP(t)-159-Rev | ATTAGATACTCCAAACGCAACTAACTTCGGGTTCA |
| Seq-pksP(p)-126-For | GCACTTAAAGACTCCTCTTTTCAA |
| Seq-pksP-6606_For | CGGGACAGAAGGCGAA |
| crRNA sequences | |
| cr5′pksP | GCCGCGCAAACCACTCGCCA |
| cr3′pksP | GTGGGGTGAGTTCCTAGGTT |
a
Underlined sequences are the microhomology arms; bold sequences are for the amplification of the HygR cassette.