TABLE 2.
List of primers used in this study
| Primer | 5′–3′ sequencea |
|---|---|
| Upstream_PilF.Deletion.F1 | CTTTGGTCGATAACATTGCG |
| Upstream_PilF.Deletion.R1 | CCTATTTTTTGTGAATCG GCAATGAAGATTGCCTTCAG |
| ErmC_PilF.Deletion.F2 | GAAGGCAATCTTCATTGC CGATTCACAAAAAATAGGCACAC |
| ErmC_PilF.Deletion.R2 | CACAGACAGGAGAAACAA ATGAACGAGAAAAATATAAAACACAGTC |
| Downstream_PilF.Deletion.F3 | GACTGTGTTTTATATTTTTCTCGTTCAT TTGTTTCTCCTGTCTGTG |
| Downstream_PilF.Deletion.R3 | AAGCTGAGTACATTGTGC |
| SK36_PheS.F1 | GCTGGAACTACTGTCTTTCT ATGACGAAAACGATTGAAGA |
| SK36_PheS.R1 | AGCCATTGATAATATCTCCT CTACTTAAACTGCTGAGAAAAAC |
| KanR.F2 | GCAGAAACCTTGTTCATCAA CTAAAACAATTCATCCAGTAAAA |
| KanR.R2 | AAGTAGAGGAGATATTATCA ATGGCTAAAATGAGAATATCAC |
| pDL278_Linearization.F3 | TACTGGATGAATTGTTTTAG TTGATGAACAAGGTTTCTGC |
| pDL278_Linearization.R3 | TCTTCAATCGTTTTCGTCAT AGAAAGACAGTAGTTCCAGC |
| PheS-Kan_SDM(A316G).F4 | GAGAAATACTCTGGATTTGGT TTTGGTCTCGGTCAAGAGCG |
| PheS-Kan_SDM(A316G).R4 | ACCAAATCCAGAGTATTTCTCTGCATCAATGC |
| Upstream_PilF.PheS*-Kan.F1 | CTTTGGTCGATAACATTGCG |
| Upstream_PilF.PheS*-Kan.R1 | GAATTGTTTTAGTTGATG GCAATGAAGATTGCCTTC |
| PilF::PheS*-Kan.F2 | CACAGACAGGAGAAACAA ATCCCATTATGCTTTGGCAG |
| PilF::PheS*-Kan.R2 | GAAGGCAATCTTCATTGC CATCAACTAAAACAATTC |
| Downstream_PilF.PheS*-Kan.F3 | CTGCCAAAGCATAATGGGAT TTGTTTCTCCTGTCTGTG |
| Downstream_PilF.PheS*-Kan.R3 | AAGCTGAGTACATTGTGC |
| Upstream_PilB.Deletion.F1 | AACTGAATTTTGGAGTCGTTAAGAG |
| Downstream_PilB.Deletion.R3 | CTTGGTTGAGTTGATTGTCGTAAT |
| Downstream_PilB.PheS*-Kan.R1 | TACTGGATGAATTGTTTTAG CAAGATCTTTGGTTTGTGTC |
| Upstream_PilB.PheS*-Kan.F2 | GACACAAACCAAAGATCTTG CTAAAACAATTCATCCAGTA |
| Downstream_PilB.PheS*-Kan.R2 | TTAAATAGGAAAGAATGATG ATCCCATTATGCTTTGGCAG |
| Upstream_PilB.PheS*-Kan.F3 | CTGCCAAAGCATAATGGGAT CATCATTCTTTCCTATTTAA |
| Downstream_PilB.CD.R1 | TTAAATAGGAAAGAATGATG CAAGATCTTTGGTTTGTGTC |
| Upstream_PilB.CD.F2 | GACACAAACCAAAGATCTTG CATCATTCTTTCCTATTTAA |
| Downstream_PilB.MIDAS.R1 | CAGCTGTTTCCTTCGTTTTT GCT ACA GCA GGT GCC ATGGCTTATGGATTAAGGAATG |
| Upstream_PilB.MIDAS.F2 | CATTCCTTAATCCATAAGCCAT GGC ACC TGC TGT AGC AAAAACGAAGGAAACAGCTG |
a
Segments highlighted in bold indicate the target sequence, while segments highlighted in italics indicate homologous recombination sequences. Underlined segments are the codon sequences used for site-directed mutagenesis.