TABLE 1.
Primers used for screening of pKpQIL-like plasmids in this study
| PCR | Primer no.a | Name | Sequence | Size (bp) | Target | Reference |
|---|---|---|---|---|---|---|
| PCR-I | 1 | FIIK-repA-F1 | CTTCACGTCCCGTTTTGATT | 657 | IncFII repA gene | 34 |
| 2 | FIIK-repA-R1 | CGCTTCAGCGCTTCTTTATC | 34 | |||
| PCR-II | 3 | QIL-F1 | ACAGGGAGTGCCAGGAAAG | 2,001 | Junction between Tn4401 tnpR and upstream IncFIIK2 backbone gene | This study |
| 4 | QIL-R1 | TGTATTTGCATGGCGATGAG | This study | |||
| PCR-III | 5 | Tn4401v-F(3098U) | TGACCCTGAGCGGCGAAAGC | 604b | pKpQIL-associated Tn4401a isoform | 6 |
| 6 | Tn4401v-R1 | GCAAGCCGCTCCCTCTCCAG | 35 | |||
| 7 | Tn4401v-R(3781L) | CACAGCGGCAGCAAGAAAGC | 6 | |||
| PCR-IV | 8 | QIL-F2 | GCCTCAGATAGATGCGGTAGC | 1,831 | Junction between Tn4401 ISKpn6 and downstream gene | This study |
| 9 | QIL-R2 | AAGCTGGAGACATGGAATGG | This study | |||
| PCR-V | 10 | QIL-hsdR-F1 | GGGTCGTTCACAAAGTCGAT | 498 | IncFIIK2-associated type I restriction modification system hsdR gene | This study |
| 11 | QIL-hsdR-R1 | CGTTGAGCACTTCACCAAAA | This study | |||
| PCR-VI | 12 | K2-repB-F1 | CCATTCCGATCCTTTTCTGA | 395 | IncFIIK2 repFIB gene | This study |
| 13 | K2-repB-R1 | AACGCTACTGTCCAGCCTGT | This study |
a
The locations of the primers are illustrated in Fig. 1.
b
The sizes are variable for different Tn4401 isoforms. Isoform b, 703bp, 382bp; a, 604bp; c, 487bp; d, 635bp, 314bp; e, 448bp.