TABLE 1.

E. coli strains and plasmids

NameGenotype or characteristicsSource or references
E. coli strains
    DH5αφ80lacZΔM15 recA1 endA1 gyrA96 thi-1 hsdR17(rK mK+) supE44 relA1 deoR Δ(lacZYA-argF)U169 17
    MG1655Sequenced λ and F derivative of K-12 3
    TOP10FmcrA Δ(mrr-hsdRMS-mcrBC) (φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL endA1 nupGInvitrogen
    TP8503Δ(lac-proB) leu thi-1 supE42 fhuA 12
    XL1Blue recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB lac1qZΔM15 Tn10]Stratagene
Plasmids
    pBNpBR322 Ω [EcoRI-HindIII: 79 bp, polylinker of pNUB193]This study
    pBNLCpBN Ω [SmaI-SmaI: 4,725 bp, lacZ-lacY′-cat from pRS551-SL]This study
    pBNLCPSpBNLC Ω [StuI-StuI: 242 bp, Plac promoter]This study
    pBR322Apr Tcr; cloning vectorGenBank no. J01749
    pBR325Apr Tcr Cmr; cloning vectorGenBank no. L08855
    pCP20Apr CmrrepA(Ts); pSC101-based vector expressing the Flp recombinase 5
    pKD4Apr KmroriR plasmid containing an FRT-aph-FRT cassette 7
    pKO3Cmr; pSC101-based vector; repA(Ts) with a sacB conferring sucrose sensitivity 11
    pNUB193pUC19 derivative with additional restriction sites in its polylinkerNew England BioLabs Inc.
    pRS551Apr KmrlacZ; operon fusion vector 19
    pRS551-SLSame as pRS551 but without tryptophan terminator upstream of lacZThis study
    pSLO3AprlacZ; promoter cloning vector 10
    pTOF1pUC18 Ω [EcoRI-HindIII: 162 bp, NotI-FRT-SmaI-FRT-NotI]; AprThis study
    pTOF2     and -3pTOF1 Ω [HincII-HincII: 1,252 bp, aph gene from pUC4K]; Apr KmrThis study
    pTOF20pTOF1 Ω [SmaI-SmaI: 4,967 bp, LacZ-cat-Plac of pBNLCS]; Apr CmrThis study
    pTOF24pK03 Ω [HincII-HincII: 1,252 bp, aph from pUC4K]; Cmr Kmr Ts SucsThis study
    pTOF25pKO3 Ω [BamHI-BamHI: 1,264 bp, aph from pUC4K]; Cmr Kmr Ts SucsThis study
    pTOF27Site-directed mutagenesis of pTOF20 adding a SnaBI site in lacY; Apr CmrThis study
    pTOF29     and -30pTOF27 Ω [HincII-HincII: 1,252 bp, aph gene from pUC4K]; Apr KmrThis study
    pTOF60Site directed mutagenesis of pTOF27 adding a SnaBI site upstream of Plac; Apr CmrThis study
    pTOF61Site-directed mutagenesis of pBR322 adding a NotI site in tet; AprThis study
    pTOF70pTOF61 Ω [NotI-NotI: 5,114 bp, FRT-lacZ-cat-Plac-FRT from pTOF60]; Apr CmrThis study
    pTOF72     and -73pTOF70 Ω [HincII-HincII: 1,252 bp, aph gene from pUC4K] Apr KmrThis study
    pUC4KApr Kmr; cloning vectorGenBank no. X06404
    pUC18Apr; cloning vectorGenBank no. L08752
    pUC19Apr; cloning vectorGenBank no. M77789