TABLE 1.
Strain or plasmid | Origin and/or genetic featuresa | Source or referencef |
---|---|---|
T. kodakarensis strains | ||
KW128 | ΔpyrF; ΔtrpE::pyrF | 21 |
TS372 | KW128 with trpE-PhmtB-TK1761 insertion | 18 |
TS416 | TS372 with nonsense mutation at codon 3 of TK1761 | 18 |
TS502 | KW128 with trpE-PhmtB-(His)10TK1827 insertion | This study |
TS503 | KW128 with trpE-PhmtB-TK1827 insertion with TAG at codon 5 | This study |
TS517 | KW128 with ΔTK0664b | This study |
TS521 | KW128 with ΔTK0664::trpE | This study |
TS538 | TS517 with ΔTK1761-TK1762-TK1763 | This study; Fig. S2 |
TS541 | TS517 with ΔTK1827 | This study; Fig. S2 |
TS1079 | TS541 with ΔTK1761-TK1762-TK1763 | This study |
TS559 | TS517 with ΔTK0149c | This study |
TS561 | TS517 with ΔTK0148-TK0149 | This study |
PdaD | KU216 ΔpyrF with ΔTK0148-pdaD | 6 |
Plasmids | ||
pTS502 | pUC118::PhmtB-(His)10TK1827-trpE-TK1828 | This study |
pTS503 | pUC118::PhmtB-TK1827-trpE-TK1828 with TAG at codon 5 of TK1827 | This study |
pTS517 | pUC118::TK0662-TK0663-TK0664-trpE-TK0662-TK0663-TK0665 | This study |
pTS521 | pUC118::TK0662-TK0663-trpE-TK0665 | This study |
pTS535 | pUC118 with MCS1-(trp-6MPs)-MCS2d | This study; Fig. S1 |
pTS538 | pTS535::TK1760-TK1764-(trp-6MPS)-TK1761 | This study; Fig. S2 |
pTS541 | pTS535::TK1825-TK1826-TK1828-(trp-6MPs)-TK1827-TK1828 | This study; Fig. S2 |
pTS559 | pTS535::TK0147-TK0148-TK0150-(trp-6MPs)-TK0150-TK0151 | This study |
pTS561 | pTS535::TK0147-TK0150-(trp-6MPs)-TK0150-TK0151 | This study |
pLC70 | trpE; Pgdh-PF1848e | 19 |
pTS436 | pLC70::PhmtB-TK1761 | This study |
pTS543 | pLC70::PhmtB-TK0149 | This study |
Loss of TK0664 results in resistance to 6-methyl purine (6MPr).
Loss of TK0149, or loss of TK0148 plus TK0149, results in agmatine auxotrophy.
The trp-6MPs cassette has P2279 and PhmtB directing the divergent transcription of trpE (TK0254) and TK0664, respectively. TK0664 expression results in sensitivity to 6-methyl purine (6MPs). Flanking the cassette are multiple cloning sites 1 and 2 (MCS1 and MCS2) that have single cleavage sites for HindIII, SphI, BglII, BclI, ClaI, NheI, SpeI, NotI, SacII, ApaI, AccI, AscI, EcoRV, and BstEII and for EcoRI, KpnI, Acc65I, BamHI, XbaI, respectively (see Fig. S1 in the supplemental material).
Figures S1 and S2 may be found in the supplemental material.