Supplementary Data

Table 1. Plasmids used in this study.

Plasmid / Insert locationa / Description and construction or referenceb
pGEM-3Zf / - / Vector containing SP6 and T7 promoters (Promega)
pEJ414 / - / lacZ reporter vector (29)
pEJ544 / 1-315 / upstream region of Rv2719c in pEJ414 (9)
pEJ559 / 1-315 / pEJ544 with TAC®GCG in s70-like –10 region, by sdm with oligos 2719pr-10bf and 2719pr-10br
pEJ597 / 1-315 / pEJ544 with CTA®AGC in recAP1-like -10 region, by sdm with oligos 2719pr-10ef and 2719pr-10er
pEJ615 / 1-315 / pEJ544 with TAC®GCG in s70-like –10 region and CTA®AGC in recAP1-like -10 region, by sdm of pEJ559 with oligos 2719pr-10ef and 2719pr-10er
pEJ629 / 179-248 / 70bp ds oligonucleotide from annealing 2719cproxprf and 2719cproxprr between XbaI and HindIII sites in pEJ414
pEJ630 / 138-182 / 45bp ds oligonucleotide from annealing 2719cdistprf and 2719cdistprr between XbaI and HindIII sites in pEJ414
pEJ631 / 179-248 / pEJ629 with TAC®GCG in s70-like –10 region, by sdm with oligos 2719pr-10bf and 2719pr-10br
pEJ632 / 138-182 / pEJ630 with CTA®AGC in recAP1-like -10 region, by sdm with oligos 2719pr-10ef and 2719pr-10er
pEJ639 / 1-315 / upstream region of Rv2719c in pGEM-3Zf, from PCR with Rv2719cfXba and Rv2719crHind

a Insert locations according to the sequence in 5, with the first base of that sequence being 1.

b ds = double-stranded, sdm = site-directed mutagenesis


Table 2. Oligonucleotides.

Name / Sequence /

Usea

Rv2719cf / CGATTACTCTCTGGCTTGGTTTG / Taqman primer
Rv2719cr / CCGAACCGCCGGTGAT / Taqman primer
Rv2719cp / CATCTGCCCGAACTGCGCCA, 5’FAM-3’TAMRA-labelled / Taqman probe
sigAf / CCGATCTCGTTGGACCAGA / Taqman primer
sigAr / CCTCGCTGTCTTCGATGAAAT / Taqman primer
sigAp / AGCTGGCTGTCGCCCTCGTCG, 5’FAM-3’TAMRA-labelled / Taqman probe
XbaRv2719c / GGTCTAGAATTGAGCGGATCGGGGGTATGC / Primer extension
LACR / TTCCCAGTCACGACGTTGTAAAA / Primer extension
Rv2719cfXba / CGTCTAGAGAGGTGTCGTTGCTGTCGTTCA / PCR for cloning
Rv2719crHind / CGAAGCTTATTGAGCGGATCGGGGGTAT / PCR for cloning
2719pr-10bf / GAACACATGAGTGACGCGAGTGTCGAACACGCG / Mutagensis
2719pr-10br / CGCGTGTTCGACACTCGCGTCACTCATGTGTTC / Mutagensis
2719pr-10ef / GTCGGTGGTGTGGTAGCCCGTTTTGCGCGTG / Mutagensis
2719pr-10er / CACGCGCAAAACGGGCTACCACACCACCGAC / Mutagensis
2719cproxprf / CTAGCGACACGCTTCGATCAAATGTTCGGATATCGAA
CACATGAGTGACTACAGTGTCGAACACGCGAGCGAT / Cloning
2719cproxprr / AGCTATCGCTCGCGTGTTCGACACTGTAGTCACTCAT
GTGTTCGATATCCGAACATTTGATCGAAGCGTGTCG / Cloning
2719cdistprf / CTAGAACCGAAACGTGTCGGTGGTGTGGTCTACCGTT
TTGCGCGTGCG / Cloning
2719cdistprr / AGCTCGCACGCGCAAAACGGTAGACCACACCACCGA
CACGTTTCGGTT / Cloning

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