Table 1 Primers used in this study.
From: An integrative expression vector for Actinosynnema pretiosum
Primer | Sequence (5' – 3') |
|---|---|
Vector construction | |
   IS117-KpnI | TGACTGGTACCGGAGTCGGGGATGTTCTTGTTC* |
   IS117-NheI | TGACTGCTAGCGTACCGGTGCCCCGATAGAC |
   ermE-XbaI/KpnI | AAAATCTAGAGGTACCAGCCCGACCCGAGCA |
   ermE-BamHI/EcoRV | AAAAGATATCGGATCCACCAACCGGCACGA |
   xylE-F4Bam | CGCGGATCCAGCGATGAACAAAGGTGTAATGCGACC |
   xylE-FEBam | CGCGGATCCAAGCTAACGTAAGGAGGAAAAACATATGAACAAAGGTGTAATGCGACCGGGCCAT |
   xylE-FABam | CGAGGATCCTCCCCACAGACAGGGAGCCCAGCATATGAACAAAGGTGTAATGCGACCGGGCCAT |
   xylE-REcoRI | GCGGAATTCTCAGGTCAGCACGGTC |
   rifH-FNdeI | CGCATACATATGGTGAAGCGGCAGCCGGACTTCG |
   rifH-RASE | CGCGAATTCACTAGTCCTAGGCTAGCGCAGCATCTTCGCGATCG |
   xylE-FSpeI | AATACTAGTATGAACAAAGGTGTAATGCGACCGGGCCAT |
   xylE-FRBSSpeI | CGCAATACTAGTAAGCTAACGTAAGGAGGAAAAACATATGAACAAAGGTGTAATGCGACCGGGCCAT |
qPCR | |
   rifH-668F | AGATCTACGTCAGCCACGAAATG |
   rifH-788R | TCGCCGATCCACAGGAA |
   16S-F | CAGAAGAAGCACCGGCTAAC |
   16S-R | TTAAGCCCCAAGTTTTCACG |
Southern hybridization | |
   IS117-F | CTGAACTCACCGCGACGTATCG |
   IS117-R | GCTAGCGTACCGGTGCCCCG |
Plasmid rescue and sequencing | |
   intseq1 | CCGTTTGGCCTCCGACTAAC |
   intseq2 | GCTCTATCGGGAGGCCTCAC |
   attM5'1 | GGAGCAGACGCTCGTCCGGG |
   siteA-F | GTCGTGGTCACCAGGCGGAC |
   siteA-R | ACCGCCTTCGTCCGTGCCAG |
   siteB-F | GATCTGTGTTCGCGGAGAGC |
   siteB-R | ACAAGGGCGTCAGCGAGGAA |
   siteC-F | GGTGTCCTCGATGCGGAAGA |
   siteC-R | AAGTCCCCGTGCTTCTCCAG |
   siteD-F | GACCCCCTCCGCGAACTGGT |
   siteD-R | GGCCTGCGAGGTTTGTGCGC |