TABLE 1.
Plasmids
| Plasmid | Antibiotic selection | Descriptiona | Size (bp) | Source, reference, or construction strategy |
|---|---|---|---|---|
| pCR2.1 | Km, Am | Cloning vector | 3,900 | Invitrogen |
| pCR-blunt | Km | Cloning vector | 3,500 | Invitrogen |
| pET29c(+) | Km | Expression vector | 5,372 | Novagen |
| pUC19 | Am | Cloning vector | 2,686 | Bioline |
| pSU18 | Cm | Cloning vector | 2,300 | 2 |
| pSU4601 | Km | ColE1::kan | 7,930 | 5 |
| R64drd-11 | Tc, Sm | R64 derepressed for transfer | 56,700 | 20 |
| pUIV201 | Am | pUC19::oriT(ColE1) | 2,886 | 38 |
| pUIV206 | Km | pET29c::mbeA | 6,974 | 38 |
| pUIV230 | Cm, Mob+ | pSU18::mob(ColE1) | 4,700 | 38 |
| pUIV235 | Km, Am | pCR2.1::mbeC* | 4,239 | This study; the 339-bp PCR fragment using the primers mobC-plus and mobC-minus cloned at pCR2.1 |
| pUIV236 | Km | pET29c::mbeC* | 5,591 | This study; the 317-bp NdeI-BamHI fragment from pUIV235 cloned at the corresponding sites of pET29c |
| pUIV238 | Km, Am | pCR2.1::mbeC† | 4,268 | This study; the 368-bp PCR fragment using the primers mobC-P1-plus and mobC-minus cloned at pCR2.1 |
| pUIV239 | Km | pET29c::mbeC† | 5,615 | This study; the 341-bp NdeI-BamHI fragment from pUIV238 cloned at the corresponding sites of pET29c |
| pUIV241 | Km, Am | pCR2.1::89-bp oriT(ColE1) | 4,016 | This study; the 116-bp PCR fragment using the primers plus-oriT and minus-oriT cloned at pCR2.1 |
| pUIV244 | Km | pCR-blunt::(1389-1855)ColE1 | 3,967 | This study; the 467-bp PCR fragment using the primers plus-ΔmbeC and minus-ΔmbeC cloned at pCR-blunt |
| pUIV245 | Cm | pSU18::89-bp oriT(ColE1) | 2,338 | This study; the 89-bp EcoRI-HindIII fragment from pUIV241 cloned at the corresponding sites of pSU18 |
| pUIV247 | Km | pET29c::mob(ColE1)ΔmbeC | 7,267 | This study; the 446-bp NdeI-XbaI fragment from pUIV244 cloned at the corresponding sites of pUIV206 |
| pUIV248 | Cm | pSU18::mob(ColE1)ΔmbeC | 4,310 | This study; the 2,037-bp XbaI-EcoRI fragment from pUIV247 cloned at the corresponding sites of pSU18 |
| pUIV261 | Km | pCR-blunt::mbeC(R13A) | 3,866 | This study; the 366-bp PCR fragment using the primers mbeC-mutR and mobC-minus cloned at pCR-blunt |
| pUIV262 | Km | pET29c::mbeC(R13A) | 5,615 | This study; the 341-bp NdeI-BamHI fragment from pUIV261 cloned at the corresponding sites of pET29c |
| pUIV264 | Km | pCR-blunt::mob(ColE1)ΔoriT | 5,441 | This study; the 1,941-bp PCR fragment using the primers mbeC-P1-plus and minus-ab cloned at pCR-blunt |
| pUIV265 | Cm | pSU18::mob(ColE1)ΔoriT | 4,201 | This study; the 1,941-bp BamHI-EcoRI fragment from pUIV264 cloned at the corresponding sites of pSU18 |
| pUIV266 | Km | pET29c::mob(ColE1)ΔoriT | 7,215 | This study; the 1,941-bp NdeI-BamHI fragment from pUIV264 cloned at the corresponding sites of pET29c |
| pUIV267 | Km | pCR-blunt::oriT(ColE1)ΔIR | 3,549 | This study; hybridizing the ΔIR-plus and ΔIR-minus primers and cloning the dsDNA in pCR-blunt |
a
*, mbeC cloned from the second ATG (coordinates 1867 to 1869); †, mbeC cloned from the first ATG (coordinates 1843 to 1845).