Supplemental Table S1.1. Oligonucleotides Used in This Study

Santi McKinney, 2014

Supplemental Table S1.1. Oligonucleotides used in this study.

Oligo Name / Sequence 5'–3'
parB_UpF / CCTTAATTAACGCACCAGATCGGTGATCAC
parB_UpR / CCTAGGAAGTCGGTGTCAGCGGACGA
parB_DnF / CCTAGGAATATTCTCGTTCTGGGCGCTCATCA
parB_DnR / TTGGCGCGCCTTCGGTCTCATGCAGCCGAT
smc_KIN_UpF / CCTTAATTAAGAACAGGTGACCGCGCCCGCGC
smc_KIN_UpR / GGAATATTACCTGAGCCGCTGGGGTTGGAGACCAGCT
dnaX_UpF / CCTTAATTAAGGATGACCAGCGGATACACG
dnaX_UpR / GGAATATTGCTCTCTACCGCAAGTACCGCCC
dnaX_DnF / GGAATATTCACGGGTGAAGGCTACCGAT
dnaX_DnR / CCGCTCGAGCGACCATCAAGGGCATATGG
mch_F / GATATCGTGAGCAAGGGCGAGGAGGA
mch_R / GATATCACCTGAGCCCTTGTACAGCTCGTCCATGC
gfp_F / GATATCGCTAGCAAAGGAGAAGAACTTTTC
gfp-R / GATATCTTTGTAGAGCTCATCCATGCC
ssb_F / CTAGCTAGCGCTGGTGACACCACCATC
ssb_R / GCTAGCACCTGAGCCGAAGGGCGGTTC
parBko_UpF / ccttaattaaCGCACCAGATCGGTGATCAC
parBko_UpR / cctaggAAGTCGGTGTCAGCGGACGA
parBko_DnF / CCTAGGGCTCGTCTCCTGCTCATCAC
parBko_DnR / GGCGCGCCCGAACGCTCGTCATCGATCT
smc_UpF / CCTTAATTAAAGCTCATGAAGATCGCACTG
smc_UpR / GGAATATTCACGACGCCTTACCCTACCGC
smc_DnF / GGAATATTTGAGCCCCGGCCGACCCCCG
smc_DnR / GCTCGAGCAGACGGCCCTCTGCGGGCG
gfp_F1 / GATATCGCTAGCAAAGGAGAAGAACTTTTC
gfp_R1 / GATATCACCTGAGCCTTTGTAGAGCTCATCCATGC
dnaN_seq_UpF / GTGAGCTCACCGACCTGTCG
lacI_3R / ATGTGTAATCCCAGCAGC
gfp_seq2F / GGGTGAAGGTGATGCTACATAC
dnaN_seq_DnR / AAATCCACCGGGAGGTCTTC
parB_seq_UpF / AGTCCCCCAACACCTTCA
parB_seq_UpR / ACAACGAGGACTACACCA
parB_seq_DnF / TGGTGTAGTCCTCGTTGT
parB_seq_DnR / GGGATGACGATCCTCAAT
smckin_seq_UpF / AACTCGCCACCGAACGGCAGCTG
smc_seq_DnR / CTCGGTGCGCACGTTCTTGGC
dnaX_seq_UpF / GGCCTGCAGCACGATCAGGT
dnaX_seq_DnR / GAGATGATGCACGACTCCGT
smc_seq_UpF / TACGCCAGTCTATCCCCAGCA
smc_seq_UpR / CGGTCGATCGTCTTCTTGGGC
smc_seq_DnF / CACCGTCCTGCCGAAATGACC
parB_ko_seq_DnR / CGACATGGGTGACCGACGCG
gfp_F2 / AAAACTGCAGGCTAGCAAAGGAGAAGAACTTTTC
gfp_R2 / CCGGAATTCTTATTTGTAGAGCTCATCCATGCC
tetR_F / CGCGGATCCGGTGTCTAGATTAGATAAAAGTAAAG
tetR_R / AAAACTGCAGGATGTCAGACCCACTTTCACAT

Restriction enzyme sites are underlined.

Supplemental Table S1.2. Plasmids used in this study.

Plasmid Name / Description / Source
pCR2.1-TOPO / ApR, KmR. PCR cloning vector. / Invitrogen
pMV261 / KmR.hsp60 promoter. / [2]
pJG1100 / KmR, HygR, sacB. Allelic-exchange suicide vector
containing aph, hyg,and sacB markers. / J. Gomez
pND235 / KmR. pMV361-based vector containing GFP under the
control of the UV15A promoter. / [2]
pND239 / HygR. pMV361-based vector containing DsRed2 under
the control of the UV15A promoter. / N. Dhar
pIS220 / KmR. pMV361 containing wag31-gfp under the control
of the hsp60 promoter. / [1]
pIS222 / KmR, HygR, sacB. pJG1100::gfp_dnaN translational
fusion for allelic exchange. / This study
pIS219 / KmR, HygR, sacB. pJG1100::gfp_dnaX translational
fusion for allelic exchange. / This study
pIS251 / KmR, HygR, sacB. pJG1100::parB_mCherry
translational fusion for allelic exchange. / This study
pIS272 / KmR, HygR, sacB. pJG1100::smc_gfp translational
fusion for allelic exchange. / This study
pIS243 / KmR. pMV235-based vector containing ssb_gfp under
the control of the UV15A promoter. / This study
pIS294 / KmR, HygR, sacB. pJG1100 ∆parB to delete parB gene
by allelic exchange. / This study
pIS239 / KmR, HygR, sacB. pJG1100 ∆smc to delete smc gene
by allelic exchange. / This study
pIS280 / GnR, HygR, pND239-based vector containing 256-tetO
repeats and tetR-gfp under control of hsp60 promoter. / This study
pBS34 / KmR, pFA6-GFPkan based vector containing mCherry / YRC, UW
pLAU44 / GnR, AmpR, 256 copies of tetO with Gm in the middle / [3]
pLAU53 / AmpR, pBAD24-lacICFP-tetR-YFP / [3]

Abbreviations: Ap, ampicillin; GFP, green fluorescent protein; Hyg, hygromycin; Km, kanamycin; YRC, UW, Yeast Research Center, University of Washington.

Supplemental Table S1.3. Bacterial strains used in this study.

Strains / Description / Source
mCherry-dnaN / mCherry-dnaN replaces dnaN in the chromosome / [1]
mCherry-dnaN pPHsp60_wag31-gfp / mCherry-dnaN replaces dnaN in the chromosome
pIS220: hsp60 promoter driving Wag31-GFP / [1]
mCherry-dnaN
pPUV_ssb-gfp / mCherry-dnaN replaces dnaN in the chromosome
pIS243: UV15A promoter driving SSB-GFP / This study
mCherry-dnaN
gfp-dnaX / mCherry-dnaN replaces dnaN in the chromosome
gfp-dnaX replaces dnaX in the chromosome / This study
mCherry-dnaN
pFROS-attB / mCherry-dnaN strain carrying the plasmid pIS280
pIS280: FROS-attB system (tetO array + tetR-gfp) / This study
parB-mCherry / parB-mCherry replaces parB in the chromosome / This study
parB-mCherrypPUV_ssb-gfp / parB-mCherry replaces parB in the chromosome
pIS243: UV15A promoter driving SSB-GFP / This study
parB-mCherry dnaN-gfp / parB-mCherry replaces parB in the chromosome
gfp-dnaN replaces dnaN in the chromosome / This study
WT
pPUV_ssb-gfp / wild-type background
pIS243: UV15A promoter driving SSB-GFP / This study
WT
pFROS-attB / wild-type background
pIS280: FROS-attB system (tetO array + tetR-gfp) / This study
mCherry-dnaN
pFROS-attB / mCherry-dnaN replaces dnaN in the chromosome
pIS280: FROS-attB system (tetO array + tetR-gfp) / This study
∆parB / in-frame deletion of chromosomalparB gene / This study
∆parB
mCherry-dnaN / in-frame deletion of chromosomalparB gene
mCherry-dnaN replaces dnaN in the chromosome / This study
∆parB
mCherry-dnaN
pPHsp60_wag31-gfp / in-frame deletion of chromosomalparB gene
mCherry-dnaN replaces dnaN in the chromosome
pIS220: hsp60 promoter driving Wag31-GFP / This study
∆parB
pFROS-attB / in-frame deletion of chromosomalparB gene
pIS280: FROS-attB system (tetO array + tetR-gfp) / This study
∆smc / in-frame deletion of chromosomalsmc gene / This study
∆smc
mCherry-dnaN / in-frame deletion of chromosomalsmc gene
mCherry-dnaN replaces dnaN in the chromosome / This study
∆smc
mCherry-dnaN
pPHsp60_wag31-gfp / in-frame deletion of chromosomalsmc gene
mCherry-dnaN replaces dnaN in the chromosome
pIS220: hsp60 promoter driving Wag31-GFP / This study
∆smc
pFROS-attB / in-frame deletion of chromosomalsmc gene
pIS280: FROS-attB system (tetO array + tetR-gfp) / This study
∆smc
parB-mCherry / in-frame deletion of chromosomalsmc gene
parB-mCherry replaces parB in the chromosome / This study
smc-gfp / smc-gfpreplaces smc in the chromosome / This study
mCherry-dnaN
smc-gfp / mCherry-dnaNreplaces dnaN in the chromosome
smc-gfpreplaces smc in the chromosome / This study
parB-mCherry
smc-gfp / parB-mCherry replaces parB in the chromosome
smc-gfpreplaces smc in the chromosome / This study
∆parB
∆smc / in-frame deletion of chromosomalparB gene
in-frame deletion of chromosomalsmc gene / This study
WT
pND235 / wild-type background
pND235: UV15A promoter driving GFP / This study

1