Table 1.

Oligonucleotides used in this study

PrimerSequence (5′-3′)aDescription
ON1 GGCCATGATATC ATTCTTGAAGACGAAAGGGCCTCG EcoRV; 5′ end of ColE1 ori-bla
ON2 GGCCATGATATC CTGATATTGTCTGCATTTGCGCCG EcoRV; 3′ end of ColE1 ori-bla
ON3 GGCCATATCGAT CCTGATATTGTCTGCATTTGCGCCG ClaI; 5′ end of lacA-front
ON4 GGCCATATCGAT TCACAGTGGCAATCTCCCCCGTAT ClaI; 3′ end of lacA-front
ON5 GGCCATATCGAT TTCAAGCTATATTTGGAGTTGAGC ClaI; 5′ end of lacA-back
ON6 GGCCATCCCGGGCTAATGTGTGTTTACGACAATTCT SmaI; 3′ end oflacA-back
ON7 GGCCATCCGCGG GAGCTC GGATCCCATTTCCCCCTTTGATTT SacII andSacI; 5′ end of xylose cassette
ON8 GGCCATCCCGGG GAGCTC CTAACTTATAGGGGTAACACTTAA SmaI and SacI; 3′ end of xylose cassette
ON9 GGCCATCCCGGGGACGTTCTTGCCATTGCTGCATAAA SmaI; 5′ end oft 0
ON10 GGCCATATCGAT ATCTCTGCAGTCGCGATGATTAAT ClaI: 3′ end of t 0
ON11 GGCCATATCGAT GCGGCCGC TCTAGAGCAACGTTCTTGCCATTG SacII andNotI; 5′ end of erm
ON12 GGCCATCTCGAG GCGGCCGC ACTCTTCCTTTTTCAATATTATTG ClaI and NotI; 3′ end of erm
ON13 GGCCATATCGATTTGACATTTTTCTTGTGGATCTGTATAATAAAGAATAATTA ClaI and −35 and −10 regions of a hybrid promoter; 5′ end of spec
ON14 GGCCATATCGATCAATAGTTACAAATTGTTTCACTA ClaI; 3′ end of spec
ON15 GGCCATGAGCTCAGGCCTTACACAGCCCAGTCCAGACTATTCGGCA SacI; Plac expression cassette
ON16 GGCCATGAGCTCAGGCCTTAACTCACATTAATTGCGTTGCGCTCACTA SacI; Plac expression cassette
ON17 GGCCATGGATCCATGAATGTGTTATCCTCAATTTGTTAC BamHI; 5′ end ofbgaB
ON18 GGCCATGGATCCCTAAACCTTCCCGGCTTCATCATG BamHI; 3′ end of bgaB
ON19 GGCCATGTCGACCTAAATTTTAACTTAATTTATAATTAAACG SalI; 5′ end ofbgaB
ON20 GGCCATGCATGCCTAAACCTTCCCGGCTTCATCATG SphI; 3′ end of bgaB
ON21 GGCCATGCATGCATGTTAACAAATCGTCAGCTGCT BamHI: 5′ end of hrcA
ON22 GGCCATGCATGCTGCCAAAATTCCCTTATTCATCATACAG BamHI; 3′ end ofhrcA
  • a G/C clamps are shown in italics; restriction sites are underlined.