Skip to main content

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