TABLE 1.
Variant promoter/operator sequences used in this study
| Variant | Test plasmid(s)a | Sequenceb | Oligonucleotides |
|---|---|---|---|
| −120 truncation | pKJ217/218 | ggatccAATTCGGCATGCTTAACGATTATTCTCTAAAATCCTAACGAATGGTGGCATGCACCAGAATCGAGGCCACTTTGAGGCTTGGAATATCCTCTCGGGGGGGATAGCATCAAGGGCAtctaga | oKJ527/564 (PCR) |
| −39 truncation | pKJ219/220 | ggatccGGCTTGGAATATCCTCTCGGGGGGGATAGCATCAAGGGCAtctaga | oKJ498/499 |
| −30 truncation | pKJ221/222 | ggatccTATCCTCTCGGGGGGGATAGCATCAAGGGCAtctaga | oKJ575/576 |
| −39LOF | pKJ225/226 | ggatccGGCaacGAATATCCTCTCGGGGGGGATAGCATCAAGGGCAtctaga | oKJ502/503 |
| −39GOF | pKJ227/228 | ggatccGGCTTGaAATATCCTCTCGGGGGGGATAtaATCAAGGGCAtctaga | oKJ504/505 |
| o_mut1 | pKJ238 | ggatccGGCTTGaAATAaaaTCTCGGGGGGGATAtaATCAAGGGCAtctaga | oKJ554/555 |
| o_mut2 | pKJ240 | ggatccGGCTTGaAATATCCaaaCGGGGGGGATAtaATCAAGGGCAtctaga | oKJ556/557 |
| o_mut3 | pKJ242 | ggatccGGCTTGaAATATCCTCTaaaGGGGGATAtaATCAAGGGCAtctaga | oKJ558/559 |
| o_mut4 | pKJ244 | ggatccGGCTTGaAATATCCTCTCGGaaaGGATAtaATCAAGGGCAtctaga | oKJ560/561 |
| o_mut5 | pKJ246 | ggatccGGCTTGaAATATCCTCTCGGGGGaaATAtaATCAAGGGCAtctaga | oKJ562/563 |
| o_mut6 | pKJ231 | ggatccGGCTTGaAATATCCTCTCGGGGaGGATAtaATCAAGGGCAtctaga | oKJ565/566 |
| o_mut7 | pKJ232 | ggatccGGCTTGaAATATCCTCTCcaGGGGGATAtaATCAAGGGCAtctaga | oKJ567/568 |
| o_mut8 | pKJ233 | ggatccGGCTTGaAATATCCTCTCcaGGaGGATAtaATCAAGGGCAtctaga | oKJ569/570 |
| o_mut9 | pKJ234 | ggatccGGCTTGaAATATCCcCTCGGGGaGGATAtaATCAAGGGCAtctaga | oKJ571/572 |
| o_mut10 | pKJ235 | ggatccGGCTTGaAATATCCcCTCcaGGGGGATAtaATCAAGGGCAtctaga | oKJ573/574 |
a
Plasmids are compatible for use in A. fabrum or E. coli. Strains harboring these plasmids are listed in Table S1. Where two plasmids are listed, the first is nreA−, and the second is nreA+. If only one plasmid is listed, it is nreA+.
b
Sequence includes the BamHI (GGATCC) and XbaI (TCTAGA) sites used for ligating into pKJ138 (nreA−) and/or pKJ140 (nreA+). Lowercase letters within the sequence represent cloning sites. Bold lowercase letters represent variations compared to the −39 truncation clone. For empty vectors pKJ138 and pKJ140, the BamHI/XbaI region contains the following sequence: ggatccACCtctaga.