Table 2. Plasmids constructed during this work.
| Plasmid | Relevant features |
| pAKE600 derivatives | |
| pKGB6.67 | parA67 inserted as EcoRI/SalI fragment |
| pKLB60.4 | PCR-amplified parA1–47 with the use of primers 1 and 6, inserted as EcoRI/HindIII fragment |
| pKLB60.5 | parAΔ48–59, PCR-amplified fragment (primers 8 and 2) encoding parA60–262 cloned as HindIII/SalI fragment into pKLB60.4 |
| pKLB60.6 | PCR-amplified parA1–76 with the use of primers 1 and 7, inserted as EcoRI/HindIII fragment |
| pKLB60.7 | parAΔ77–85, PCR-amplified fragment (primers 9 and 2) encoding parA86–262 cloned as HindIII/SalI fragment into pKLB60.6 |
| pKLB60.8* | parA84 inserted as EcoRI/SalI fragment |
| pUC18 derivatives | |
| pGMB11* | parA74 inserted as EcoRI/SalI fragment |
| pGMB12* | parA75 inserted as EcoRI/SalI fragment |
| pGMB13* | parA76 inserted as EcoRI/SalI fragment |
| pGMB14* | parA78 inserted as EcoRI/SalI fragment |
| pGMB15* | parA79 inserted as EcoRI/SalI fragment |
| pBGS18 derivatives | |
| pGMB33* | parA inserted as EcoRI/SalI fragment with internal SacI site without amino acid change |
| Bacterial two-hybrid vectors | |
| pKGB4 | pUT18 derivative with modified MCS to facilitate ORF cloning as EcoRI/KpnI or EcoRI/SacI fragments in-frame with the N terminus of CyaT18 |
| pKGB4.14 | pKGB4 with parA67–cyaT18 translational fusion |
| pKGB4.15 | pKGB4 with parA68–cyaT18 translational fusion |
| pKGB4.16 | pKGB4 with parA70–cyaT18 translational fusion |
| pKGB4.17 | pKGB4 with parA72–cyaT18 translational fusion |
| pKGB4.18 | pKGB4 with parA74–cyaT18 translational fusion |
| pKGB4.19 | pKGB4 with parA75–cyaT18 translational fusion |
| pKGB4.20 | pKGB4 with parA76–cyaT18 translational fusion |
| pKGB4.21 | pKGB4 with parA78–cyaT18 translational fusion |
| pKGB4.22 | pKGB4 with parA79–cyaT18 translational fusion |
| pKGB4.24 | pKGB4 with parA82–cyaT18 translational fusion |
| pKGB4.25 | pKGB4 with parA83–cyaT18 translational fusion |
| pKGB4.26 | pKGB4 with parA84a†–cyaT18 translational fusion |
| pKGB4.27 | pKGB4 with parA84–cyaT18 translational fusion |
| pKGB4.28 | pKGB4 with parA85–cyaT18 translational fusion |
| pKGB5 | pKNT25 derivative with modified MCS to facilitate ORF cloning as EcoRI/KpnI or EcoRI/SacI fragments in-frame with the N terminus of CyaT25 |
| pKGB5.14 | pKGB5 with parA67–cyaT25 translational fusion |
| pKGB5.15 | pKGB5 with parA68–cyaT25 translational fusion |
| pKGB5.16 | pKGB5 with parA70–cyaT25 translational fusion |
| pKGB5.17 | pKGB5 with parA72–cyaT25 translational fusion |
| pKGB5.18 | pKGB5 with parA74–cyaT25 translational fusion |
| pKGB5.19 | pKGB5 with parA75–cyaT25 translational fusion |
| pKGB5.20 | pKGB5 with parA76–cyaT25 translational fusion |
| pKGB5.21 | pKGB5 with parA78–cyaT25 translational fusion |
| pKGB5.22 | pKGB5 with parA79–cyaT25 translational fusion |
| pKGB5.24 | pKGB5 with parA82–cyaT25 translational fusion |
| pKGB5.25 | pKGB5 with parA83–cyaT25 translational fusion |
| pKGB5.26 | pKGB5 with parA84a–cyaT25 translational fusion |
| pKGB5.27 | pKGB5 with parA84–cyaT25 translational fusion |
| pKGB5.28 | pKGB5 with parA85–cyaT25 translational fusion |
| pBBRMCS-1 derivative | |
| pABB1.0 | pBBRMCS-1 with modified EcoRI restriction site within the CmR cassette (PCR site-directed mutagenesis with primers 26 and 27) |
| pABB1.2 | pAMB9.37 with tacp–parB transcriptional fusion |
| pABB84 | pAMB9.37 with tacp–parA84a transcriptional fusion |
| pET28a (+) or pET28mod derivatives | |
| pABB8.0 | parA fragment PCR-amplified with primers 3 and 4 and inserted as NcoI/XhoI |
| pABB8.67 | parA67 PCR-amplified and inserted as above |
| pABB8.78 | parA78 PCR-amplified and inserted as above |
| pABB8.83 | parA83 PCR-amplified and inserted as above |
| pABB8.84 | parA84a PCR-amplified and inserted as above |
| pKLB8.3 | parA40–262 inserted as EcoRI/SalI fragment |
| pKLB8.4 | parA1–40 inserted as EcoRI/SalI fragment |
| pKLB8.5 | parA1–151 inserted as EcoRI/SalI fragment |
| pKLB8.6 | parA152–262 inserted as EcoRI/SalI fragment |
| pKLB8.7 | parA40–151 inserted as EcoRI/SalI fragment |
| pKLB8.8 | parA1–85 inserted as EcoRI/SalI fragment |
| pKLB8.9 | parA86–151 inserted as EcoRI/SalI fragment |
| pKLB8.10 | parAΔ48–59 inserted as EcoRI/SalI fragment |
| pKLB8.11 | parAΔ77–85 inserted as EcoRI/SalI fragment |
| Yeast two-hybrid vectors | |
| pABB482* | pGMB57 with parA82 inserted as EcoRI/SalI fragment |
| pABB483* | pGMB57 with parA83 inserted as EcoRI/SalI fragment |
| pABB484* | pGMB57 with parA84a inserted as EcoRI/SalI fragment |
| pABB485* | pGMB57 with parA85 inserted as EcoRI/SalI fragment |
| pGMB21* | pGAD424 with parA67 inserted as EcoRI/SalI fragment |
| pGMB22* | pGAD424 with parA68 inserted as EcoRI/SalI fragment |
| pGMB24* | pGAD424 with parA70 inserted as EcoRI/SalI fragment |
| pGMB25* | pGAD424 with parA72 inserted as EcoRI/SalI fragment |
| pGMB57 | pGAD424 with gal4AD–parA40–262 fragment with internal SacI site inserted as BamHI/SalI fragment |
| pKLB4.5 | pGAD424 with gal4AD–parA1–151 inserted as EcoRI/SalI fragment |
| pKLB4.8 | pKLB4.5 with gal4AD–parA1–85 with SalI oligo with stop codon inserted in NaeI site |
| pKLB4.9 | pKLB4.5 with gal4AD–parA86–151 with EcoRI oligo with ATG codon inserted in NaeI site, EcoRI fragment deleted |
| pKLB6.3 | pBTM116 with lexADB–parA40–262 inserted as BamHI/SalI fragment |
| pKLB6.4 | pBTM116 with lexADB–parA1–40 inserted as EcoRI/SalI fragment |
| pKLB6.5 | pBTM116 with lexADB–parA1–151 inserted as EcoRI/SalI fragment |
| pKLB6.6 | pBTM116 with lexADB–parA152–262 inserted as EcoRI/SalI fragment |
| pKLB6.7 | pBTM116 with lexADB–parA40–151 inserted as EcoRI/SalI fragment |
| pKLB6.8 | pBTM116 with lexADB–parA1–85 inserted as EcoRI/SalI fragment |
| pKLB6.9 | pBTM116 with lexADB–parA86–151 inserted as EcoRI/SalI fragment |
| pKLB6.10 | pBTM116 with lexADB–parAΔ48–59 inserted as EcoRI/SalI fragment |
| pKLB6.11 | pBTM116 with lexADB–parAΔ77–85 inserted as EcoRI/SalI fragment |
*
Construction of these plasmids is described in detail in Supplementary Materials and Methods.
†
parA84a encodes ParAL84A, whereas parA84 encodes ParAL84K.