Table 3 Primers used in this study
From: A Streptomyces coelicolor host for the heterologous expression of Type III polyketide synthase genes
Primer | Sequence 5′–3′ | Description |
|---|---|---|
pIJ86F1 | ACGCCTGGTCGATGTCGGAC | Sequencing primers for recombinant plasmids of pIJ86 and pIJ12477 |
pIJ86R2 | TGCGGTCAGTGCGTGTGTCG | |
SCO1206F1 | ATCCCCAAGACCGAGGACTG | Sequencing primer for internal sequence of sco1206-1207-1208 |
SCO1206F2 | AGGCCGTCATGGACCGCCAG | |
SCO1206F3 | TGTTCCACCTGCTGCTGAGC | |
SCO1206F4 | AGTGGTGAGCGGCCTGGTC | |
SCO1206FBglII | AAAAAAGATCTCGCAAGCCTTCCGCGAGGCG | PCR of sco1206-1207-1208 for cloning into pIJ12477 |
SCO1208RHindIII | AAAAAAAGCTTCTAACCGTGTCGCAGCGGCGTGAG | |
SCO1206FSpe | CGCAAGCCTTCCGCGAGGCGAAAGCAGGCACGGTTCATGACTAGTATTCCGGGGATCCGTCGACC | Flanking primers for sco1206-1207-1208 disruption cassette |
SCO1208RNhe | GATCGCGCGGACGCTGGCCGGGATGCCGATCCGCTAACCGCTAGCTGTAGGCTGGAGCTGCTTC | |
SCO1206TF | TCGAGCTGGCCAAGCTG | Test primers for verification of AprR cassette replacements and in-frame deleted regions |
SCO1208TR | TGGAGTACGCGCAGACC | |
SCO7221TF | AGTTGCATGGGTCACTGC | |
SCO7221TR | ATGTACCGGCCCAGAGC | |
SCO7671TF | GTTGCACCAGCCGATGG | |
SCO7669TR | GTGAGTCGATGACTGTCGTGG | |
SCO7221F1 | AGTCGGTGCTCCGGCTGGAC | Sequencing primer for internal sequence of sco7221 |
SCO7221FBamHI | AAAAAGGATCCCCTCACCTGCTCCGCAGCAGACCC | PCR of sco7221 for cloning into pIJ86 and pIJ12477 |
SCO7221RHindIII | AAAAAAAGCTTCTACAGCCACTCCCCTTCCAGAGCGGTGG | |
SCO7221FSpe | CGCCGTGATACACGGCGAGCACTCCGTCGAGACCCGAGAACTAGTATTCCGGGGATCCGTCGACC | Flanking primers for sco7221 disruption cassette |
SCO7221RNhe | GTGGCGGTCAGGCCCGGGGCCGCCGCGTACAGGACGGCAGCTAGCTGTAGGCTGGAGCTGCTTC | |
SCO7671FSpe | GTACGCACGGTAGTGGGGCCGGCGGCCGAGGAAGGCATGACTAGTATTCCGGGGATCCGTCGACC | Flanking primers for sco7669-7670-7671 disruption cassette |
SCO7669RNhe | GTCCGCTACGCGCGCGCCGCGCGGGGCCGGAACGGCTCAGCTAGCTGTAGGCTGGAGCTGCTTC | |
SVEN5367F1 | GGCCCGACACCGAGGACTG | Sequencing primer for internal sequence of sven5367 |
SVEN5367FBamHI | AAAAAGGATCCCGGGTGAGTGTGGGCGGCAGTTC | PCR of sven5367 for cloning into pIJ12477 |
SVEN5367RHindIII | AAAAAAAGCTTTCAGGCGACGGACGTGCGGG |