## 4 Materials and Methods
### Bacterial Strains, Plasmids and Media
\begin{table}
\begin{tabular}{l l c} \hline \hline
**Strains** & **Genotype** & **Reference** \\ \hline BW25113 & _lac_I_ \(rmB_{14}\) \(\Delta lacZ_{\rm W16}\) _lsndR514_ & [70] \\  & _\(\Delta araBAD_{\rm AH33}\) \(\Delta rhBaAD_{\rm LD78}\)_ & [19] \\ GK1942 & BW25124\(\pm\)PDK06 & [19] \\ W3110 & \(\lambda^{-}\), _IN (rmD-rmE)I, rph-1_ & CGSC, Yale \\ GK6075 & BW25113 _lapC190\(<\)c\(\rightarrow\)_ & [6] \\ SR23529 & GK6075 _lapC190\(<\)jrt_ & This study \\ SR23583 & BW25113 _lapC190\(<\)aph_ & This study \\ SR23679 & BW25113 _lapC_\(>\)aph_ & [28] \\ SR23743 & SR23679 _lapC_\(>\)_eff_ & [28] \\ SR9073 & BW25113 _mlc_\(<\)aph_ & [42] \\ SR9078 & BW25113 _mlaD\(>\)aph_ & This study \\ SR23627 & BW25113 _mlaD\(<\)aph_ & This study \\ SR23630 & GK6075 _mlaD_\(<\)aph_ & This study \\ SR19957 & BW25113 _srrA\(<\)aph_ & [36] \\ SR23605 & BW25113 _marmA\(<\)aph_ & This study \\ SR27092 & BW25113 _clsA\(<\)aph_ & Our collection \\ SR23320 & BW25113 _clsA\(<\)jrt_ & [28] \\ SR20066 & BW25113 _dsA\(<\)aph_ & [36] \\ SR24224 & BW25113 _pldA\(<\)aph_ & This study \\ GK1275 & W3110 _lpxM\(<\)aph_ & [13] \\ GK1077 & W3110 _lpxL\(<\)aph_ & [13] \\ SR23941 & GK6075 _lpxL\(<\)aph mshA_ 1412P & This study \\ SR23946 & GK6075 _lpxM\(<\)aph mshA_ 1412P & This study \\ SR19761 & BW25113 + PCA24N & This study \\ SR23951 & SR23529 + psrrA\({}^{+}\) & This study \\ SR23952 & SR23951 _clsA\(:\)_Tn10_ & This study \\ SR23992 & SR23529 + ppf/\(\delta M^{+}\) & This study \\ SR23994 & SR23529 + ppf/\(\delta M^{+}\) & This study \\ SR23996 & SR23529 + ppf/\(\delta M^{+}\) & This study \\ SR23998 & SR23529 + pexp\({}^{+}\) & This study \\ SR24000 & SR23529 + paccB\({}^{+}\) & This study \\ SR24027 & SR23529 + plapD\({}^{+}\) & This study \\ SR24097 & SR23529 + ppldA\({}^{+}\) & This study \\ SR23660 & SR23591 + pmarA\({}^{+}\) & This study \\ SR24099 & SR23529 + pmarA\({}^{+}\) _mlaC\(<\)aph_ & This study \\ SR24100 & SR23529 + pmarA\({}^{+}\) _mlaC\(:\)_Tn10_ & This study \\ SR24101 & SR23529 + pgs\(A^{+}\) & This study \\ SR24103 & SR23529 + pycJ\({}^{+}\) & This study \\ SR24105 & SR23529 + pac\(pT^{+}\) & This study \\ SR24107 & SR23529 + pymg\({}^{+}\)c\({}^{+}\) & This study \\ CR3592 & BW25113 _sPhC21 _and220_:Tn10 _\(\Delta\)ftsH3\(:\)_Kan_ & [19] \\ SR24021 & SR23529 + pdfsA\({}^{+}\) & This study \\ SR24011 & SR23529 + pdfsA D74N & This study \\ SR24013 & SR23529 + pdfsA F82Y & This study \\ SR24015 & SR23529 + pdfsA S83A & This study \\ SR24017 & SR23529 + pdfsA L84A & This study \\ SR24019 & SR23529 + pdfsA E85A & This study \\ SR24056 & SR23529 + plapD \(\Delta\)N20 & This study \\ SR24058 & SR23529 + plapD \(\Delta\)(100-132) & This study \\ SR24167 & SR23529 + plapD D69A Y70A R71A & This study \\ SR24170 & SR23529 + plapD L84A L85A P86A & This study \\ SR24173 & SR23529 + plapD N93A P94A F95A & This study \\ \hline
**Plasmids** & **Genotype** & **Reference** \\ \hline pCA24N & IPTG-inducible expression vector cm\({}^{\rm R}\) & [33] \\ pDUET & Expression vector & Our collection \\ pKD3 & _oriRGKg, bla_(Amp\({}^{\rm R}\)), _kan, rgnB_(Ter), _cat_ & [70] \\ pKD13 & _oriRGKg, bla_(Amp\({}^{\rm R}\)), _kan, rgnB_(Ter) & [70] \\ pKD46 & _araBp-gam-bet-exo, bla_(Amp\({}^{\rm R}\)), _repA10_(ts) & [70] \\ \hline \hline \end{tabular}
\end{table} 
Table 6: Bacterial strains and plasmids used in this study. Identification of Multicopy Suppressors Whose Overexpression Suppresses the Temperature-Sensitive Phenotype of lapC190 Bacteria  
As _E. coli_ K-12 strains carrying the chromosomal _lapC190_ mutation exhibit the Ts phenotype (inability to grow above 42 \({}^{\circ}\)C), we used such a growth defect to identify genes whose overexpression can allow growth under such non-permissive conditions. To achieve this, competent cells of SR23529 and SR23583 _lapC190_ derivatives of BW25113 were transformed with a complete genomic library of all predicted ORFs of _E. coli_ cloned in pCA24N (ASKA collection). Transformants were selected at 43 \({}^{\circ}\)C in the presence of IPTG (75 \(\upmu\)M).