Table 2 Bacterial strains, plasmids and oligonucleotides used in this study

E. coli OmniMAX

mcrA Δ(mrr hsdRMS-mcrBC); resistant to T1 and T5 phage; used for cloning

Invitrogen

B. subtilis 1012

leuA8 metB5 trpC2 hsrM1

[22]

Plasmid

Description

Source/reference

pHCMC04-bgaB

PxylA-bgaB

[9]

pHCMC05-bgaB

Pspac-bgaB

[9]

pHT01

Pgrac01 (previously called Pgrac)

[16]

pHT01-bgaB

Pgrac01-bgaB

[16]

pHT10-gfp+

Pgrac-gfp-Strep

[16]

pHT100

Pgrac100-bgaB

[11]

pHT100-gfp+

Pgrac100-gfp

This study

pHT212

Pgrac212-bgaB

[17]

pHT253

Pgrac100-8xHis-MCS (Start codon-His-tag-BamHI-XbaI-AatII-SmaI)

This study

pHT254

Pgrac100-MCS-His (BamHI-start codon-XbaI-AatII-His-tag-stop codon/TAA)

This study

pHT255

Pgrac100-MCS-Strep (BamHI-start codon-XbaI-AatII-Strep-tag-stop codon/TAA)

This study

pHT1169

Pgrac100-His-gfp

This study

pHT1170

Pgrac100-gfp-His

From Nguyen H. N

pHT1171

Pgrac100-gfp-Strep

This study

pHT1178

Pgrac100-8xHis-bgaB

From Nguyen H. N

pHT1179

Pgrac100-bgaB-His

This study

pHT1180

Pgrac100-bgaB-Strep

This study

^a Oligonucleotide

Sequence 5′ → 3′

Used for

ON301F

GATCTATGGAAGCTCATCACCATCACCATCACCATCACGGATCCATGTCTAGAGTCGACGT

pHT253

ON302R

CGACTCTAGACATGGATCCGTGATGGTGATGGTGATGGTGATGAGCTTCCATA

pHT253

ON303F

GATCCATGTCTAGAGTCGACGTCGCTCATCACCATCACCATCACCATCACTAACGT

pHT254

ON304R

TAGTGATGGTGATGGTGATGGTGATGAGCGACGTCGACTCTAGACATG

pHT254

ON305F

GATCCATGTCTAGAGTCGACGTCGCTTGGAGCCATCCGCAATTTGAAAAATAACGT

pHT255

ON306R

TATTTTTCAAATTGCGGATGGCTCCAAGCGACGTCGACTCTAGACATG

pHT255

ON941

AAAGGAGGAAGGATCCATGAATGTGTTATC

pHT1179 and pHT1180

ON1250

CTGCCCCGGGGACGTCAACCTTCCCGGCTTCATCATGC

pHT1179 and pHT1180

ON1277

AAAGGAGGAAGGATCCATGGCTAGCAAAGGAGAAGAACT

pHT1169 and pHT1171

ON1278

GGCCATGACGTCTTTGTAAAGCTCATCCATGCCATGTGT

pHT1171

ON1279

CCAGGTCTCAGATCTATGGCTAGCAAAGGAGAAGAACT

pHT100-gfp+

ON1280

GGCCATGACGTCTTATTTGTAAAGCTCATCCATGCCATGTGT

pHT100-gfp + and pHT1169