Characterization of xiamenmycin biosynthesis reveals a
advertisement
Table S2. Primers used for constructing and confirming the mutants and for protein expression Primer Gene Sequence(5‘-3‘) For screening library ORF5317-For ORF5317-Rev ORF5310-For ORF5310-Rev ORF5317 ORF5310 GCGTGCGGACATGCGTGTTGTGGTG GGGTCCAACCTACCCAAATGTGATGGAGCAA GCGGCTGGAGTGTAGCGAGTCTGGAATG GGGTATGTCCACCCGATGCTCCTGTCTCAT For PCR Targeting TORF5311-For TORF5311-Rev TORF5313-For TORF5313-Rev TORF5314-For TORF5314-Rev TORF5315-For TORF5315-Rev TORF5316-For TORF5316-Rev ximA ximB ximC ximD ximE ATGAGACAGGAGCATCGGGTGGACATACCCGAGAACTTGTGGTTCATGTGCAGCTCCATC TCACGTTCGAGGCGCATTCGACGCCGGATAGTGACGATG TGAGCTCAGCCAATCGACTG GTGATCGATATTTCCGCTCAACCCTCGCAGCAGAGCACGTGGTTCATGTGCAGCTCCATC TCAAAAGACTCTCCCCGCAACGATGGCGACGAGCACGAGTGAGCTCAGCCAATCGACTG GTGCGCACGGAGTCGCGCAGCCTGGCCCAGTTCGTGGCGTGGTTCATGTGCAGCTCCATC TCATGCGTCGTGGACGGCGTCTCGATCGAGGAGACACGGTGAGCTCAGCCAATCGACTG ATGCCGAACTCTCCCGCCGCGGTCTTCGAGCGGCTCACCTGGTTCATGTGCAGCTCCATC TCACGTCGTCTCCATCATCGTGTACTCCTGCCGGATCCGTGAGCTCAGCCAATCGACTG ATGGGCCAGACGACGCACACAGCACTCGACCGCTACATGTGGTTCATGTGCAGCTCCATC TCAGCCCGGCGTACGGGTGTACCGGTTGCGCAGGTTCGTTGAGCTCAGCCAATCGACTG For gene verification VORF5311-For VORF5311-Rev VORF5313-For VORF5313-Rev VORF5314-For VORF5314-Rev VORF5315-For VORF5315-Rev VORF5316-For ximA ximB ximC ximD ximE GCCCGAACCCCTGAGACTCTGGAGATC GCCGCCTTGTCCTAACGGTGTTTGCC CCGAGGTGGGCTTTCTGCAAGAGTCA GCACGGACCAGGGGCCTGCAT CTGCCGGGAAAAGTTCATGGCAAACCG TGCCGAGACGAACCTCGATGCGGAC GCCGTCCACGACGCATGAGGGATC GGTCGGCGAGCTCCATGTAGCGGTC CGGCAGGAGTACACGATGATGGAGACG 1 VORF5316-Rev TCTCGGCAGCTACCGGCGCATC For heterologous expression HORF5311-6-For HORF5311-6-Rev ximA~ximE GCTCTAGACGGCTGGAGTGTAGCGAGTCTGGAATG GGAATTCCCCGGACGTGGGAGCGATAGGG For protein expression ximA-F-NdeⅠ ximA ximA-R-XhoⅠ ximB-F-NdeⅠ ximB ximB-R-XhoⅠ ximC-F-NdeⅠ ximC ximC-R-HindⅢ ximD-F-EcoRⅠ ximD ximD-R-HindⅢ ximE-F-NdeⅠ ximE ximE-R- HindⅢ TCAGCcatATGAGACAGGAGCATCGGGTGG TCAGctcgagTCACGTTCGAGGCGCATTC ctagcataTGGTGATCGATATTTCCGCTC atcgctcgagTCAAAAGACTCTCCCCGCAAC ataccataTGCGCACGGAGTCG atacaagcttTCATGCGTCGTGGACG TCAGgaattcATGCCGAACTCTCCCGC TCAGaagcttTCACGTCGTCTCCATCATCG TCGAcatATGGGCCAGACGACGCACAC TCAGaagcttTCAGCCCGGCGTACGGGT For protein coexpression ximD-F-EcoRⅠ ximD ximD-R-HindⅢ CoximE-F-NdeⅠ ximE CoximE-R-KpnⅠ TCAGgaattcATGCCGAACTCTCCCGC TCAGaagcttTCACGTCGTCTCCATCATCG TCGAcatATGGGCCAGACGACGCACAC TAGCggtaccTCAGCCCGGCGTACGGGT For RT-PCR ORF1467 ORF1467 ORF2290 ORF2290 ORF2781 ORF2781 GCCAACGCCCCCTCACCTC GCCGACCACCATCATGCTC GCGTCGGCGAAGTCGTTGA GCGTCCGTCTGCCTCTACC ACCTCGGCACCCGCACCAC 2 CGGCCACCAGTCCCACTCC ORF4925 ORF4925 ACCGTCACCCCGCACAAGT GCCATCCAGCAGAAGAACA ORF5065 ORF5065 ORF5313 ORF5313 ACGTGGTGATGTTCGAGCA GTCAGGGAGGCGGTGTAGG TGGCTGGGGATGGTCTTCG CCTTGTCCTGGTGGGCGTA 3
