Table 2.
Plasmids used in this study.
| Plasmid | Description a | Reference or Source |
|---|---|---|
| pRL60 | E.coli-Amycolatopsis shuttle plasmid, Ery R, Kan R, amy+, oriT | [29] |
| pOSV236 | E. coli-Streptomyces shuttle plasmid expressing the Xis and Int proteins for site-specific excision of excisable cassettes, Amp R, Pur R, oriT | [36] |
| pOSV504 | Source of the excisable hygromycin cassette (att1Ωhyg), Amp R, Hyg R | [25] |
| pOJ260 | E.coli-Amycolatopsis shuttle plasmid, suicide vector in Amycolatopsis, Apr R, oriT | [37] |
| pOSV805 | E.coli-Amycolatopsis shuttle plasmid, integrative in Amycolatopsis using the ϕBT1 SSR system, Hyg R, oriT | [32] |
| pCR-blunt | E. coli cloning vector, Kan R | Invitrogen |
| pT-atts | pUC derivative containing the attL and attR minimal sites from pSAM2, Amp R, Constructed by Twist Biosciences | This study |
| pRL60∆amy | E.coli-Amycolatopsis shuttle plasmid, derivative of pRL60, amy−, Ery R, Kan R, oriT | This study |
| pEA01 | E.coli-Amycolatopsis shuttle plasmid, pRL60∆amy derivative containing the xis and int genes from pSAM2 under the control of trcp, the pac gene and oriT, Ery R, Kan R, Pur R, oriT | This study |
| pEA02 | E.coli-Amycolatopsis shuttle plasmid, suicide vector in Amycolatopsis; pOJ260 derivative containing the minimal attL sequence and a multi-cloning site, Apr R, oriT | This study |
| pEA03 | E.coli-Amycolatopsis shuttle plasmid, suicide vector in Amycolatopsis; pOSV805 derivative containing the minimal attR sequence and a multi-cloning site, Hyg R, oriT | This study |
| pRIF01 | pCR-blunt derivative containing the downstream homologous region of rifK, KanR | This study |
| pRIF02 | pCR-blunt derivative containing the upstream homologous region of rifK, KanR | This study |
| pRIF05 | pOJ260 derivative containing the excisable hygromycin cassette (att1Ωhyg) flanked by the upstream and downstream homologous region of rifK, for replacement of rifK, Apr R, oriT | This study |
| pRIF09 | pCR-blunt derivative containing the downstream homologous region of the rif cluster, Kan R | This study |
| pRIF10 | pCR-blunt derivative containing the upstream homologous region of the rif cluster, Kan R | This study |
| pRIF12 | E.coli-Amycolatopsis shuttle plasmid, suicide plasmid in Amycolatopsis; pEA02 derivative containing the upstream region of the rif cluster, attL, Apr R, oriT | This study |
| pRIF14 | E.coli-Amycolatopsis shuttle plasmid, suicide plasmid in Amycolatopsis; pEA03 derivative containing the downstream region of the rif cluster, attR, Hyg R, oriT | This study |
a Abbreviations: amy—α-amylase; R—Resistance, Ery—Erythromycin, Kan—Kanamycin, Amp—Ampicillin, Pur—Puromycin, Apr—Apramycin, Hyg—Hygromycin.