Table S2
Amplification target
Primers 5’-3’
Primer
name
Strain
bcwcl1
upstream non-coding region
bcwcl1
upstream non-coding region
bcwcl1
downstream non-coding region
bcwcl1
downstream non-coding region
hph resistance cassette (from
pLOB1 vector)
hph resistance cassette (from
pLOB1 vector)
bcwcl1
upstream non-coding region
GCGGATAACAATTTCACACAGGAAACAGCACCAGCTGTGGAAGTGTCTG
(FW)
CGAATCGGGAATGCGGCTCCACAGCTGCAGGGTATTTGAGCGGAGGCAGG
(RC)
TTAATGATGATTACTAACAGATATCAAGCTTTGTCATGTTGTTATAGTCTCGG
(FW)
GTAACGCCAGGGTTTTCCCAGTCACGACGATCTGTATCGGGGTTGTGTTT
(RC)
CTGCAGCTGTGGAGCCGCATT(FW)
LC-256
AAGCTTGATATCTGTTAGTAA(RC)
LC-270
GCGGATAACAATTTCACACAGGAAACAGCAGGATATACCGAGGGACGAG
(FW)
oligo1
bcwcl1
upstream non-coding region
GACCGGGATCCACTTAACGTTACTGAAATCATTGAAGAAATTACTCATGATA
(RC)
oligo2
bcwcl1
downstream non-coding region
CAAAAATGCTCCTTCAATATCATCTTCTGTCTTGGCAGGGTTCTCCTGGAT
(FW)
oligo3
bcwcl1
downstream non-coding region
GTAACGCCAGGGTTTTCCCAGTCACGACGGTTGAATTGGAAAACTTGGA
(RC)
oligo4
hph resistance cassette (from
pCSN44 vector)
GACAGAAGATGATATTGAAGGAGC (FW)
oL768
hph resistance cassette (from
pCSN44 vector)
GATTTCAGTAACGTTAAGTGG (RC)
oL687
Amplification
of
genetic
constructs (from pRS426)
Amplification
of
genetic
constructs (from pRS426)
Amplification of bcwcl1, to be
fused to PoliC promoter
GGCAGTGAGCGCAACGCAAT (FW)
oL83
∆bcwcl1
mutant 1
∆bcwcl1
mutant 1
∆bcwcl1
mutant 1
∆bcwcl1
mutant 1
∆bcwcl1
mutant 1
∆bcwcl1
mutant 1
∆bcwcl1
mutants 2
& 3
∆bcwcl1
mutants 2
& 3
∆bcwcl1
mutants 2
& 3
∆bcwcl1
mutants 2
& 3
∆bcwcl1
mutants 2
& 3
∆bcwcl1
mutants 2
& 3
-
ATTCAGGCTGCGCAACTGTT (RC)
oL84
-
TCCATCACATCACAATCGATCCAACCATGCCAATGACCCAAGCAGACTTG
(FW)
Amplification of bcwcl1, to be
fused to Tgluc terminator
AATCATACATCTTATCTACATACGCTAACCCTCCTTGAAAATCTCATTA
(RC)
5’
integration,
at
locus; diagnostic PCR
bcwcl1
GAGAGACAGGATGGTACAAATGAG (FW)
bcwcl1
PoliCF
bcwcl1
TglucR
oL588
3’
integration,
at
locus; diagnostic PCR
bcwcl1
ACACTTGTGCTTGTGCTTGC (RC)
oL589
5’
integration,
at
PoliC
promoter; diagnostic PCR
3’
integration,
at
“tubA”
terminator; diagnostic PCR
5’ integration, at hph CDS;
diagnostic PCR
GGTACTGCCCCACTTAGTGGCAGCTCGCG (RC)
oL585
GGTCCTCGGAGTGCAGATGGG (FW)
oL584
ATGGCTGTGTAGAAGTACTC (FW)
oL32
3’ integration,
diagnostic PCR
TCGCCCTCCGAGAGCTGCAT (RC)
oL29
GTATCAACCCCCACGGCTCTCA (FW)
CGGCGAATGCGGAATACCA (RC)
GGCATCTCTTGGAGGAAGAA (FW)
oL586
oL587
oL1226
CGCATATCAGCATATCGAGATGTCC (FW)
oL1716
at
hph
CDS;
bcwcl1 ORF, diagnostic PCR
bcwcl1 ORF, diagnostic PCR
bcwcl1
5’
integration
at
bcniaD locus, diagnostic PCR
5’
integration
at
bcniaD
locus, diagnostic PCR
LC-276
LC-277
LC-278
LC-269
∆bcwcl1
+bcwcl1
∆bcwcl1
+bcwcl1
∆bcwcl1
mutant 1,
2 & 3
∆bcwcl1
mutant 1,
2 & 3
∆bcwcl1
mutant 1
∆bcwcl1
mutant 1
∆bcwcl1
mutants 2
& 3
∆bcwcl1
mutants 2
& 3
∆bcwcl1
+bcwcl1
∆bcwcl1
+bcwcl1