Table 2 Plasmids used in this study

pSB1A3

bla, ori pMB1, rfp

Registry of Standard Biological Parts

pSB1A3-P_xylA

bla, ori pMB1, P_xylA (xylose inducible promoter)

[52]

pBS2E

bla, ori ColE1, lacA^’, MLS^r, rfp, ‘lacA

[52]

pBS1C

bla, ori ColE1, amyE’, rfp, cat, ‘amyE

[52]

pBS1Z

bla, ori ColE1, amyE’, zeo^r, ‘amyE

Modification of pBS1C replacing the chloramphenicol with a zeocin resistance marker

pXT

bla, ori ColE1, thrC’, P_xylA, spec^r, ‘thrC, MLS^r

[59]

pDG780

Source of kan^r

[60]

pBS1C-P_xylA

bla, ori ColE1, amyE’, cat^r, P_xylA, ‘amyE

P_xylA excised from pBS1A3-P_xylA and cloned into pBS1C3 using EcoRI and SpeI

pBS2E-P_xylA

bla, ori ColE1, lacA^’, MLS^r, P_xylA, ‘lacA

P_xylA excised from pBS1A3-P_xylA and cloned into pBS2E using EcoRI and SpeI

pBS2E-P_xylA-ureABCEFGD^Sp

bla, ori ColE1, lacA^’, MLS^r, P_xylA, ureABCEFGD^Sp, ‘lacA

ureABCEFGD amplified from S. pasteurii DSM33 gDNA using primers SG0508/SG0509 and cloned into pBS2E-P_xylA using SpeI

pXT-P_xylA-ureABCEFGD^Bp

ori, bla, thrC’, P_xylA, ureABCEFGD^Bp, spec^r, ‘thrC, MLS^r

ureABCEFGD amplified from B. paralicheniformis ATCC9945a gDNA using primers SG0652/SG0653. PCR fragment cut with BsmBI and cloned into pXT cut with BamHI and EcoRI

pSB1A3-UT^Bp

bla, ori pMB1, UT^Bp

UT-family Urease transporter amplified from 9945a gDNA using primers SG0646/SG0647 and cloned into pSB1A3 using EcoRI and PstI

pSB1A3-ureH^Bp

bla, ori pMB1, ureH^Bp

ureH amplified from B. paralicheniformis ATCC9945a gDNA using primers SG0648/SG0649 and cloned into pSB1A3 using XbaI and PstI. The plasmid then underwent QuikChange PCR to remove the EcoRI site in ureH using primers SG0650/SG0651

pBS1C-P_xylA-UT^Bp

bla, ori ColE1, amyE’, cat, P_xylA, UT^Bp ‘amyE

UT-family urease transporter excised from pSB1A3-UT^Bp with XbaI and SpeI and cloned into pBS1C-P_xylA cut with SpeI and PstI

pBS1C-P_xylA-ureH^Bp

bla, ori ColE1, amyE’, cat, P_xylA, ureH^Bp ‘amyE

ureH excised from pSB1A3-ureH^Bp with XbaI and SpeI and cloned into pBS1C-P_xylA cut with SpeI and PstI

pBS1C-P_xylA-UT^Bp-ureH^Bp

bla, ori ColE1, amyE’, cat, P_xylA, UT^Bp, ureH^Bp ‘amyE

ureH excised from pSB1A3-ureH^Bp with XbaI and SpeI and cloned into pBS1C-P_xylA-UT^Bp cut with SpeI and PstI