Table 3 Bacterial strains, plasmids and primers used in this study

Escherichia coli

TOP10F'

F' {lacI^q, Tn10(Tet^R)} mcrA Δ(mrr-hsdRMS-mcrBC) f80dlacZ DM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu) 7697 galU galK rpsL (Str^R) endA1 nupG

Invitrogen

Pseudomonas aeruginosa

PAO1

Wild-type

[47]

PKM400

PAOampP::aacCI; Gm^R

This study

PKM500

PAOampG::aacCI; Gm^R

This study

PKM300

PAOampR::aacCI; Gm^R

[10]

PKM301

PAOattB::P _ampC -lacZ; Tc^R

[10]

PKM303

PAOampR::aacCI attB::P _ampC -lacZ; Tc^RGm^R

[10]

PKM104

PAOattB::P _ampP -lacZ; Tc^R

KKF0290, This study

PKM105

PAOattB::P _ampG -lacZ; Tc^R

This study

PKM312

PAOampR::aacCI attB::P _ampG -lacZ; Tc^RGm^R

This study

PKM313

PAOampR::aacCI attB::P _ampP -lacZ; Tc^RGm^R

This study

PKM404

PAOampP::aacCI attB::P _ampP -lacZ; Tc^RGm^R

This study

PKM405

PAOampP::aacCI attB::P _ampG -lacZ; Tc^RGm^R

This study

PKM504

PAOampG::aacCI attB::P _ampP -lacZ; Tc^RGm^R

This study

PKM505

PAOampG::aacCI attB::P _ampG -lacZ; Tc^RGm^R

This study

PKM506

PAOampG::aaCI attB::P _ampC -lacZ; Tc^RGm^R

This study

PKM507

PAOampP::aaCI attB::P _ampC -lacZ; Tc^RGm^R

This study

Plasmids

pCRII-TOPO

Ap^R, Km^R; ColE1ori lacZα

Invitrogen

pUCGm

Ap^R, Gm^R; pUC19 derivative containing gentamycin cassette

[38]

pEX100T

Ap^R; sacB oriT

[39]

pMF54

ColE1-SF broad-host replicon

[48]

pME6030

Tc^R; oriV_pVS1 oriV_p15A oriT

[41]

pRK2013

Km^R; ColE1ori-Tra (RK2)⁺

[40]

pTrcphoA

Ap^R; low-copy trc promoter expression vector carrying the lacI^q and phoA

[43]

pTrclacZ

Ap^R, low-copy trc promoter expression vector carrying the lacI^q and lacZ

[43]

pSJ10

Tc^R; CTX-lacZ fused with ampC promoter, P _ampC

[10]

pKKF003

Ap^R, Km^R; pCRII-TOPO with a 2904-bp fragment containing PAO1 coordinates 4921591-4924494 (PA4392-PA4393/ampF-ampG)

This study

pKKF004

Ap^R, Km^R; pCRII-TOPO with a 2779-bp fragment containing PAO1 coordinates 4721496-4724275 (PA4217-PA4218/ampO-ampP)

This study

pKKF069

Tc^R; pME6030 with a 2904-bp Eco RI flanked fragment containing ampF-ampG

This study

pKKF073

Tc^R; pME6030 with a 2779-bp Eco RI flanked fragment containing ampO-ampP

This study

pKKF087

Tc^R; CTX-lacZ fused with ampP promoter, P _ampP

This study

pKKF091

Tc^R; CTX-lacZ fused with ampG promoter, P _ampG

This study

pKKF145

Ap^R, Gm^R; pCRII-TOPO derivative with ampP::aacCI

This study

pKKF149

Ap^R, Gm^R; pCRII-TOPO derivative with ampG::aacCI

This study

pKKF157

Ap^R, Gm^R; pEX100T derivative with ampG::aacCI

This study

pKKF161

Ap^R, Gm^R; pEX100T derivative with ampP::aacCI

This study

pAA0115

Ap^R, pTrcphoA with 1.9 kb PstI DNA from pMF54 containing stabilization fragment

This study

pAA0112

Ap^R, Km^R; pCRII-TOPO containing a 1797 ampG PCR product amplified using primers KKF09 and KKF10

This study

pAA0121

Ap^R, pAA0115 containing a 1,813 Eco RI fragment containing ampG from pAA0112

This study

pAA1261

Ap^R; pAA0121 digested with Bam HI and Sal I, treated with Klenow and re-ligated to remove an Xba I site. Used as basis for Erase-a-base system

This study

pKKF259

Ap^R; pTrcphoA derivative with a 1797-bp fragment containing PA4393

This study

pKKF458

Ap^R; pTrcphoA derivative with a 1245-bp fragment containing PA4218

This study

pKKF459

Ap^R; pTrclacZ derivative with a 1797-bp fragment containing PA4393

This study

pKKF465

Ap^R; pTrclacZ derivative with a 1245-bp fragment containing PA4218

This study

Primers

KKF01ampG For

5'-TCCAGCTTGACGTCGAGATT-3'

This study

KKF04ampG Rev

5'-AGAACATTCTCCTGGCCATGG-3'

This study

KKF05ampP For

5'-AACGGCCACGCTAGCAACAC-3'

This study

KKF08ampP Rev

5'-GTGGCGCCTGGAGTCTTG-3'

This study

KKF09ampG 2For

5'-GGGAATTCCATATGACTCAGCAATCCTGG-3'

This study

KKF10ampG 2Rev

5'-GCTCTAGATGCTCGGCGTTCTGGTGT-3'

This study

KKF13ampP 2For

5'-TCTAGATCAGGCCTCTTCCGCCCG-3'

This study

KKF14ampP 2Rev

5'-ATGCTTGAGCTGTACCGCCA-3'

This study

PA4218_9 junctionRTF

5'-ACCTCACCCTGATCCTCTG-3'

This study

PA4218_9 junctionRTR

5'-CAGGTAGAGCAACGCCAG-3'

This study

PA4392_3 junctionRTF

5'-CAACGACAGGGTGGACATAC-3'

This study

PA4392_3 junctionRTR

5'-GAGACTTGTAGGCGACCAG-3'

This study

(NS)₅RandomPrimer

5'-NSNSNSNSNS-3'

[49]