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Table 3 Plasmids used in this study

From: Knock-in/Knock-out (KIKO) vectors for rapid integration of large DNA sequences, including whole metabolic pathways, onto the Escherichia coli chromosome at well-characterised loci

Plasmid Properties Reference/ source
pKD3 oriR6Kgamma bla catP [23]
pKD4 oriR6Kgamma bla aphA [23]
pKD46 repA101ts & oriR101 ParaBexo, bet, gam araC bla [23]
pCP20 Flp recombinase expression plasmid [51]
pNPDX2 PBTac2 (Boehringer) carrying modified xynA cDNA from Neocallimastix patriciarum [80]
pBAV1K-T5-gfp pBAV1K-T5 plasmid that carries gfp gene [81]
pCSCX pCR2.1 plasmid carrying cscA, cscK and cscB gene from E. coli W [46]
pJAS01 Synthetic construct. Contains MCS & FRT-catP-FRT cassette flanked by hairpin loops. This study
parsB-R6K JSP64/JSP65 PCR product from pKD4 (PvuII; 1,495; R6K ori and bla) ΩJSP10/JSP11 PCR product (PmeI; 1,109; arsB’) This study
placZ-R6K JSP64/JSP65 PCR product from pKD4 (PvuII; 1,495; R6K ori and bla) ΩJSP57/JSP58 PCR product (PmeI; 1,318; lacZ’) This study
prbsAR-R6K JSP64/JSP65 PCR product from pKD4 (PvuII; 1,495; R6K ori and bla) ΩJSP55/JSP56 PCR product (PmeI; 1,034; rbsAR’) This study
pKIKOarsB Cm parsB-R6K (PmeI; 2,604; linear) ΩpJAS01 (PmeI; 1,143; MCS/catP cassette) This study
pKIKOlacZ Cm placZ-R6K (PmeI; 2,813; linear) ΩpJAS01 (PmeI; 1,143; MCS/catP cassette) This study
pKIKOrbsAR Cm prbsAR-R6K (PmeI; 2,529; linear) ΩpJAS01 (PmeI; 1,143; MCS/catP cassette) This study
pKIKOarsB Km pKIKOarsB Cm (XbaI; 2,839; catP drop-out) ΩpKD3 (XbaI; 930; aphA) This study
pKIKOlacZ Km pKIKOlacZ Cm (XbaI; 2,764; catP drop out) ΩpKD3 (XbaI; 930; aphA) This study
pKIKOrbsAR Km pKIKOrbsAR Cm (XbaI; 3,111; catP drop out) ΩpKD3 (XbaI; 930; aphA) This study
pKIKOarsB Ωxyn Cm pKIKOarsB Cm (PstI/SalI; 3,763) ΩpNPDX2 (PstI/SalI; 1,108; xynA) This study
pKIKOlacZ Ωxyn Cm pKIKOlacZ Cm (PstI/SalI; 3,688) ΩpNPDX2 (PstI/SalI; 1,108; xynA) This study
pKIKOrbsAR Ωxyn Cm pKIKOrbsAR Cm (PstI/SalI; 4,035) ΩpNPDX2 (PstI/SalI; 1,108; xynA) This study
pKIKOrbsAR Ωgfp Cm pKIKOrbsAR Cm (SacI/SpeI; 4,035) ΩpBAV1K-T5-gfp (Sac I/Spe I; 1,849; aphA & gfp) This study
pKIKOarsB ΩcscAKB Cm pKIKOarsB Cm (EcoRI; 3,771) ΩpCSCX (EcoRI; 4,064; cscAKB) This study