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. 2017 Jan 30;114(7):E1215–E1223. doi: 10.1073/pnas.1613609114

Table S5.

Sequences and applications of oligonucleotide primers used in this study

Primer no. Primer Sequence (5′→3′) Features
1 NMT CTCAGCCACGGTCAATGCGGCCGCGTTTTACCTGTTCTCG Forward primer used to introduce NotI restriction site
2 NMB AACAGGTAAAACGCGGCCGCATTGACCGTGGCTGAGGCTC Reverse primer used to introduce NotI restriction site
3 KMT CAACCTCGGGTAACGGTACCCTATCGCGCTTCCTGCTTTCCGG Forward primer used to introduce KpnI restriction site
4 KMB AAGCAGGAAGCGCGATAGGGTACCGTTACCCGAGGTTGCATGATAGC Reverse primer used to introduce KpnI restriction site
5 ILM264 GAAGCTCGCGGCTACCCGTGAAAAACTCACCACCACCCTCG Forward primer used to remove the non-coiled-coil region of WbbB
6 ILM265 TGGTGGTGAGTTTTTCACGGGTAGCCGCGAGCTTCTCACGTTC Reverse primer used to remove the non-coiled-coil region of WbbB
7 DW01 TCCCAAGACAAAGCGATGCGTGAGAAGCTCGCGGCTACC Forward primer used to remove four heptads from the WbbB coiled-coil (amino acids 401–434)
8 DW02 CGCGAGCTTCTCACGCATCGCTTTGTCTTGGGAAATGGC Reverse primer used to remove four heptads from the WbbB coiled-coil (amino acids 401–434)
9 DW10 gatcgctagcATGCACCATCACCATCACCATGTGAAGATACTTATTACTGGCGGG Forward primer for amplification of rmlB and cloning into pET28a(+); His-tag and NheI restriction site
10 DW11 gatcaagcttTTACTGGCGTCCTTCATAGTTCTG Reverse primer for the amplification of rmlB and cloning into pET28a(+); HindIII restriction site
11 DW12 gatcgctagcATGCACCATCACCATCACCATGTGATGATTGTGATTAAAACAGCAATACC Forward primer for amplification of rmlC and cloning into pET28a(+); His-tag and NheI restriction site
12 DW13 gatcaagcttTTACTCTGTTAACAAGGCTTGATCCAG Reverse primer for the amplification of rmlC and cloning into pET28a(+); HindIII restriction site
13 DW14 gatcgctagcATGCACCATCACCATCACCATATGAATATCTTACTTTTTGGTAAGACAG Forward primer for amplification of rmlD and cloning into pET28a(+); His-tag and NheI restriction site
14 DW15 gatcgaattcTTAGATGGTTGTCGTCGTAAACATTTC Reverse primer for the amplification of rmlD and cloning into pET28a(+); EcoRI restriction site
15 DW4 gatcgctagcATGGATTCCGCGCCGGCAGCGGCG Forward primer for the amplification of WbbB576-871 (isolation of GT1 domain) and cloning in pET28a(+); NheI restriction site
16 DW5 gatcaagctttaagtgatgtgtatggtgatgCTGAAAATACAGGTTTTCATCAATCGA CAGCAAATAACGCTTCAG Reverse primer for the amplification of WbbB576-871 (isolation of GT1 domain), introduction of a His6-tag and cloning in pET28a(+); HindIII restriction site
17 OL1028 gatcccatggGCCTGCTGTCAAAATCGAAG Forward primer for the amplification of WbbB540-1106 and cloning into pET28a(+); NcoI restriction site
18 OL1030 gatcctcgaggCGGTTGCGCTTAAACTCCG Reverse primer for the amplification of WbbB540-1106 and cloning into pET28a(+); XhoI restriction site
19 Bo/l-C-For TAAccatggaggaggtatctcatATGTCGGGCCGAATTTGC Forward primer used to introduce NcoI and NdeI restriction sites into the wbbB gene between amino acids 504 and 505
20 Bo/l-C-Rev catatgagatacctcctccatggttaGGTCAGCTTCCATGACAGGC Reverse primer used to introduce NcoI and NdeI restriction sites into the wbbB gene between amino acids 504 and 505
21 Bo/l-D-For TAAccatggaggaggtacttcatATGCAAGACAAAGCGATGAGAAAGATTTTAG Forward primer used to introduce NcoI and NdeI restriction sites into the wbbB gene between amino acids 401 and 402
22 Bo/l-D-Rev catatgaagtacctcctccatggTTAGGAAATGGCTTCATTTATCAATGATG Reverse primer used to introduce NcoI and NdeI restriction sites into the wbbB gene between amino acids 401 and 402
23 DW57 GATCCCATGGATGCTGGCTGTATTTTTACCTCCC Forward primer for the amplification of WbbBΔ402–539 (coiled-coil region), introduction a NcoI restriction site
24 DW58 gatcaagcttttacttgtcatcgtcatccttataatcGCGGTTGCGCTTAAACTCCG Reverse primer for the amplification of WbbBΔ402–539 (coiled-coil region), introduction of a FLAG tag and a HindIII restriction site
25 DW73 ATTCTACGGCGATCCTAATTGCGAA GCAGCCGCGGCGTATCTGCAGGAGC Forward primer used to mutation R738, R739, K740 to alanine.
26 DW74 GCTTCTTCAGCTCCTGCAGATACGCCGC GGC TGCTTCGCAATTAGGATCGCCG Reverse primer used to mutation R738, R739, K740 to alanine.

Restriction sites are underlined in the sequences, regions of nonchromosomal sequence are identified by lowercase, and nucleotides changed for site-directed mutations are marked in bold.