Table 2.
Plasmids used and constructed in this study
| Plasmids | Description |
|---|---|
| pTRC99A::intI1 | oriColE1 [Ap]R (Rowe-Magnus et al, 2001) |
| pSU38Δ | orip15A [Km]R (Biskri et al, 2005) |
| pSU38-attI1 | pSU38Δ::attI1 (Biskri et al, 2005) |
| pSU38-attCaadA7 | 76 bp PstI/BamHI attCaadA7 fragment (annealing between attC-GD1 and attC-GD2) in pSU38Δ digested by PstII/BamHI |
| pSU38Δ-attP | 420 bp SalI λattP fragment from pG-attP in pSU38Δ digested by SalI |
| pTSA29-CXI-AK | oriPSC101ts; intλ (Valens et al, 2004) |
| pSU711ΔoriT::aac(3)-IV | oriVR388 (IncW) [Km Gm]R (Demarre et al, 2005) |
| pG-λattP | oriColE1; λattP lacZ::λattB; [Ap]R (Valens et al, 2004) |
| pMalC-2X::intI1 | 1023 bp SmaI/BamHI intI1 PCR fragment (Fmal2 and Eibam2) amplified from pTRC99A::intI1 in pMalC-2X (New England Biolab) digested by XmnI/BamHI |
| pSW23T | pSW23T::oriTRP4; oriVR6K [Cm]R (Demarre et al, 2005) |
| pSW23TISS | 1772 bp inverse PCR fragment (Isal/sac-1 and Isal/sac-2) amplified from pSW23T, digested by BamHI and religated |
| pVCR-B | pSW23T::VCR2/1B (Biskri et al, 2005) |
| pVCR-T | 207 bp SalI/SacI VCR2/1 fragment from pVCR-B in pSW23Tiss digested by SalI/SacI |
| pAttI1-B | 155 bp EcoRI/BamHI attI1 fragment from pSU38-attI1 in pSW23T digested by EcoRI/BamHI |
| pAttI1-T | 155 bp EcoRI/BamHI attI1 fragment from pSU38-attI1 in pSW23Tiss digested by EcoRI/BamHI |
| pVCR-BIKSL | 1723 bp inverse PCR fragment (Ikpn/sal-1 and Ikpn/sal-2) amplified from pVCR-B, digested by SmaI and religated |
| pVCR-TIKSC | 1723 bp inverse PCR fragment (Ikpn/sac-1 and Ikpn/sac-2) amplified from pVCR-T, digested by SmaI and religated |
| pVCR-BINV | 260 bp KpnI/SalI oriTRP4 fragment from pVCR-B in pVCR-BIKSL digested by KpnI/SalI |
| pVCR-TINV | 260 bp KpnI/SacI oriTRP4 fragment from pVCR-T in pVCR-TIKSC digested by KpnI/SacI |
| pAttC-B | 76 bp MfeI/BamHI attCaadA7 fragment (annealing between attC-GD3 and attC-GD4) in pSW23TISS digested by EcoRI/BamHI |
| pAttC-T | 76 bp MfeI/BamHI attCaadA7 fragment (annealing between attC-GD3 and attC-GD4) in pSW23T digested by EcoRI/BamHI |
| pAttC-B-Mut1 | 70 bp EcoRI/BamHI attCaadA7-Mut1 fragment (annealing between attC-Mut1 UP and DW) in pSW23T digested by EcoRI/BamHI |
| pAttC-B-Mut2 | 70 bp EcoRI/BamHI attCaadA7-Mut2 fragment (annealing between attC-Mut2 UP and DW) in pSW23T digested by EcoRI/BamHI |
| pAttC-B-Mut3 | 70 bp EcoRI/BamHI attCaadA7-Mut3 fragment (annealing between attC-Mut1 DW and attC-Mut3) in pSW23T digested by EcoRI/BamHI |
| pAttC-B-Mut4 | 70 bp EcoRI/BamHI attCaadA7-Mut4 fragment (annealing between attC-Mut4 UP and DW) in pSW23T digested by EcoRI/BamHI |
| pλattB-1 | 34 bp EcoRI/BamHI λattB fragment (annealing between attB-1 and attB-2) in pSW23T digested by EcoRI/BamHI |
| pλattB-2 | 34 bp EcoRI/BamHI λattB fragment (annealing between attB-1 and attB-2) in pSW23TISS digested by EcoRI/BamHI |
| pSW27 | pSW23T::oriTR388; oriT orientation −; oriVR6K [Cm]R (Demarre et al, 2005) |
| pSW27ISS | 1844 bp inverse PCR fragment (Isal/sac-3 and Isal/sac-4) amplified from pSW27, digested by KpnI and religated |
| p388VCR-B | 203 bp SalI/SacI VCR2/1 fragment from pVCR-B in pSW27ISS digested by SalI/SacI |
| p388VCR-T | 184 bp EcoRI/SacI VCR2/1 fragment from pVCR-B in pSW27 digested by EcoRI/SacI |
| pΔ388VCR-B | 1678 bp inverse PCR fragment (Imfe/sac-1 and Imfe/sac-2) amplified from p388VCR-B, digested by KpnI and religated |
| pSW26 | pSW23T::oriTR388; oriT orientation +; oriVR6K [Cm]R (Demarre et al, 2005) |
| p388VCR-TINV | 184 bp EcoRI/SacI VCR2/1 fragment from p388VCR-T in pSW26 digested by EcoRI/SacI |
| p388VCR-BINV | 373 bp MfeI/SacI oriTR388 fragment from p388VCR-B in pΔ388VCR-B digested by MfeI/SacI |