TABLE 1.

Primers used for constructing pBS(Kan)ToMO, for sequencing the ToMO touABCDEF locus of P. stutzeri OX1 in pBS(Kan)ToMO and pBZ1260, and for mutagenizing the ToMO touA locus via DNA shuffling and saturation mutagenesis at codons for I100, Q141, T201, and F205

Primer	Nucleotide sequence
Construction primers
ToMO-KpnI-KACFront	5′-GGTGCGGTACCACCCATTAGCG-3′
ToMO-NotI-KACRear	5′-CTCCTGCAGCGGCCGCTGTTAATGC-3′
Sequencing primers
ToMO1	5′-GCCAAGCGCGCAATTAACCCTC-3′
ToMO2	5′-CCGTATGCCAATGTCAAGCGGAGC-3′
ToMO3	5′-CGTCAGTTGCTTGATCTAGGGCTCG-3′
ToMO4	5′-GGGTGACGACGCTCATGATTATGAG-3′
ToMO5	5′-GGGCGTTGAAGGCGGCGATATG-3′
ToMO6	5′-GGGCGGGGGATGTGGCGATTGCG-3′
ToMO7	5′-GCCCACTCAAACACGATGACTGG-3′
ToMO8	5′-GGTAGTCTTGCCCAATCCGAAGG-3′
ToMO9	5′-GCAACTCCGGCGGGTGTGGG-3′
ToMO10	5′-CGGTCTCGCCTACCTTCATCCG-3′
Mutagenesis primers
I100-front	5′-CACTATGCAACTTCACTTCGGAGCGNNNGCACTTG-3′
I100-rear	5′-GCGGCGTATTCTTCAAGTGCNNNCGCTCCGAAGTG-3′
Q141-front	5′-CCGCCATGGGCAAATCNNNCTTTAC-3′
Q141-rear	5′-GGCATACGGAAAGTAAAGNNNGATTTGCCC-3′
T201-front	5′-CGCATTCGAAACAGGCTTCNNNAATATGC-3′
T201-rear	5′-CCGAGAAACTGCATATTNNNGAAGCC-3′
F205-front	5′-GCTTCACCAATATGCAGNNNCTCGG-3′
F205-rear	5′-CAGCGGCCAAACCGAGNNNCTGC-3′
ToMO-SalI-rear	5′-CCCACTCATAATCATGAGCGTCG-3′
ToMO-KpnI-front	5′-CCGGCTCGTATGTTGTGTGGAATTGTGAGCGG-3′
ToMO-BstEII-rear	5′-CCAGGATCTTGAGCGACGGTCCACCTTGCTGTGCG-3′