Table 1 Plasmids used in this study
Designation | Relevant features | Copy number | References |
|---|---|---|---|
pBBR1MCS-1 | BHR1, IncA/C, CmR | Medium | [36] |
pBGS18 | ori MB1, KmR | High | [41] |
pET28a | ori MB1, KmR, T7p, lacO, His6- tag, T7 tag | Medium | Novagen |
pGBT30 | ori MB1, ApR, lacI q, tacp expression vector | High | [35] |
pJSB1.24 | ori MB1, KmR, TraRA3-korCp-korC (RA3 coordinates 9437-33657 nt, 3093-3705 nt) | High | [42] |
pKGB4 | pUT18 with MCS modified | High | Głąbski K.2 |
pKGB5 | pKNT25 with MCS modified | Medium | Głąbski K.2 |
pKNT25 | ori p15, KmR , lacp - MCS - cya T25 | Medium | [31] |
pKT25 | ori p15, KmR , lacp-cya T25- MCS | Medium | [31] |
pKT25-zip | ori p15, KmR , lacp-cya T25- GCN4 leucine zipper | Medium | [31] |
pLKB2 | pKT25 with MCS modified | Medium | Kusiak L.2 |
pLKB4 | pUT18C with MCS modified | High | Kusiak L.2 |
pMPB13.3 | ori RA3, KmR pABB20-lacI q -tacp-nic | Low | Przyłuski M.2 |
pMPB13.4 | ori RA3, KmR pABB20-lacI q -tacp-mobC-nic | Low | Przyłuski M.2 |
pPT01 | ori SC101, KmR, promotorless xylE | Medium | [50] |
RA3 | BHR1, IncU, CmR , SmR , SuR | Low | Hayes F.3 |
pUC18 | ori MB1, ApR | High | [23] |
pUT18 | ori ColE1, ApR , lacp- MCS-cyaT18 | High | [31] |
pUT18C | ori ColE1, ApR , lacp -cyaT18- MCS | High | [31] |
pUT18C-zip | ori ColE1, ApR , lacp -cyaT18- GCN4 leucine zipper | High | [31] |
Plasmids constructed in this study | |||
Designation | Description | ||
pJSB2.1 | pUC18-mobC, EcoRI-SalI fragment amplified by PCR with the use of primers #1 and #2 (RA3 coordinates 9837-10455 nt) | ||
pJSB2.2 | pUC18-mobC1-129, EcoRI-SalI fragment amplified by PCR with the use of primers #1 and #2 spontaneous stop codon mutation at position 10225 nt (RA3 coordinates 9837-10455 nt) | ||
pJSB2.9 | pUC18-mobCp, 417 bp SphI-BamHI fragment amplified by PCR with the use of primers #3 and #8 (RA3 coordinates 9435-9852 nt) | ||
pJSB2.11 | pUC18 with 116 bp SphI-BamHI PCR fragment amplified by PCR with the use of primers #6 and #8 (RA3 coordinates 9736-9852 nt) | ||
pJSB2.12 | pUC18 with 116 bp SphI-BamHI PCR fragment amplified by PCR with the use of primers #5 and #8 (RA3 coordinates 9736-9852 nt), mutation in a putative oriT motif | ||
pJSB2.13 | pUC18 with 100 bp SphI-BamHI PCR fragment amplified with the use of primers #5 and #4 (RA3 coordinates 9736-9836 nt), mutation in a putative oriT motif | ||
pJSB2.14 | pUC18 with 100 bp SphI-BamHI PCR fragment amplified with the use of primers #6 and #4 (RA3 coordinates 9736-9836 nt) | ||
pJSB2.15 | PCR mutagenesis of pJSB2.9 with the use of primers #9 and #10 (mutVI) | ||
pJSB2.16 | PCR mutagenesis of pJSB2.9 with the use of primers #13 and #14 (mutVII) | ||
pJSB2.17 | PCR mutagenesis of pJSB2.9 with the use of primers #11 and #12 (mutVIII) | ||
pJSB2.18 | PCR mutagenesis of pJSB2.9 with the use of primers #15 and #16 (mutIX) | ||
pJSB2.30 | pUC18 mobC1-155, EcoRI-SalI fragment amplified by PCR with the use of primers #1 and #18 carrying (RA3 coordinates 9837-10308 nt) | ||
pJSB2.43 | pUC18 with 61 bp oligonucleotides (primer #19 and #20), oligonucleotides are inserted in SmaI site in of pUC18 | ||
pJSB2.44 | pJSB2.43 digested by NheI and EcoRI, blunt-ended with the use of Klenow fragment and self-ligated, to remove IRIV | ||
pJSB2.55 | pJSB2.44 oriT45ΔOM -lacI q -tacp-mobC-nic, BamHI-SalI fragment (lacI q -tacp-mobC-nic) from pMPB13.4 | ||
pJSB2.56 | pJSB2.44 oriT45ΔOM -lacI q -tacp-nic, BamHI-SalI fragment (lacI q -tacp-nic) from pMPB13.3 | ||
pJSB2.57 | pJSB2.55 oriT45ΔOM -lacI q -tacp-mobC-nic- TraRA3-korCp-korC, SmaI-SalI fragment (TraRA3-korCp-korC) from pJSB1.24 | ||
pJSB2.58 | pJSB2.56 oriT45ΔOM -lacI q -tacp-nic- TraRA3-korCp-korC, SmaI-SalI fragment (TraRA3-korCp-korC) from pJSB1.24 | ||
pJSB4.1 | pBBR1MCS-1 lacI q, tacp-mobC, BamHI-SalI from pJSB5.1 | ||
pJSB4.2 | pBBR1MCS-1 lacI q, tacp-mobC1-129, BamHI-SalI from pJSB5.2 | ||
pJSB5.1 | pGBT30 tacp-mobC, EcoRI-SalI fragment from pJSB6.1 | ||
pJSB5.2 | pGBT30 tacp-mobC1-129, EcoRI-SalI fragment from pJSB6.2 | ||
pJSB6.1 | pET28a T7p-mobC, EcoRI-SalI fragment from pJSB2.1 | ||
pJSB6.2 | pET28a T7p-mobC1-129, EcoRI-SalI fragment from pJSB2.2, | ||
pJSB6.30 | pET28a T7p-mobC1-155, EcoRI-SalI fragment from pJSB2.30 | ||
pJSB7.9 | pPT0I mobCp-xylE, SphI-BamHI fragment from pJSB2.9 | ||
pJSB7.10 | pPT0I mobCp-xylE, 229 bp SphI-BamHI fragment PCR amplified with the use of primers #7 and #8 (RA3 coordinates 9623- 9852 nt) | ||
pJSB7.11 | pPT0I mobCp-xylE, SphI-BamHI fragment from pJSB2.11 | ||
pJSB7.12 | pPT0I mobCp-xylE, SphI-BamHI fragment from pJSB2.12 | ||
pJSB7.13 | pPT0I mobCp-xylE, SphI-BamHI fragment from pJSB2.13 | ||
pJSB7.14 | pPT0I mobCp-xylE, SphI-BamHI fragment from pJSB2.14 | ||
pJSB7.15 | pPT0I mobCp-xylE, SphI-BamHI fragment from pJSB2.15 | ||
pJSB7.16 | pPT0I mobCp-xylE, SphI-BamHI fragment from pJSB2.16 | ||
pJSB7.17 | pPT0I mobCp-xylE, SphI-BamHI fragment from pJSB2.17 | ||
pJSB7.18 | pPT0I mobCp-xylE, SphI-BamHI fragment from pJSB2.18 | ||
pJSB8.1 | pLKB4 cyaT18-mobC; EcoRI-HincII fragment from pJSB6.1 cloned between EcoRI-SmaI sites of pLKB4 | ||
pJSB8.30 | pJSB8.1 digested by ClaI and self-ligated to remove 3′ end of mobC (mobC1-155) | ||
pJSB9.1 | pKGB4 mobC-cyaT18, EcoRI-SacI fragment amplified by PCR with primers #1 and #17 | ||
pJSB10.1 | pLKB2 cyaT25-mobC; EcoRI-HincII fragment from pJSB5.1 cloned between EcoRI-SmaI sites of pLKB2 | ||
pJSB10.2 | pLKB2 cyaT25-mobC1-129; EcoRI-HincII fragment from pJSB5.2 cloned between EcoRI-SmaI sites of pLKB2 | ||
pJSB11.1 | pKGB5 mobC-cyaT25; EcoRI-SacI fragment from pJSB9.1.1 | ||