Paulsson:Plasmids

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Contents

Plasmids

  • SeqBuilder files for many plasmids can be found in the PAULSSON LAB \ DNA sequences shared folder
  • Key to drug resistance markers: Ap (ampicillin, BLA gene), Cm (chloramphenicol, CAT gene), Kn (kanamycin, APH gene(?))
  • "Ptet" and "Plac" are from Lutz and Bujard ('97) and are actually modified Plambda promoters that have tet and lac operators, and are constitutive in the absence of TetR and LacI. Names should be changed to be more acurate, as in Lutz and Bujard: PLtetO-1 and PLlacO-1

Per's plasmids

Rack I

Name Alias Marker(s) Origin Insert Notes
pPM1 Ap pSC101*ts-Rep Ptet-Citrus
pPM2 Ap pSC101*ts-Rep Plac-Citrus
pPM3 Ap pSC101*ts-Rep Ptet-Citrus-LAA (ssrA-tag)
pPM4 Ap pSC101*ts-Rep Plac-Citrus-LAA (ssrA-tag)
pPM5 Ap pSC101*ts-Rep Ptet-Citrus
pPM6 Ap pSC101*ts-Rep Plac-Citrus
pPM7 Ap pSC101*ts-Rep Ptet-Citrus-(+2)LAA (ssrA(+2)-tag)
pPM8 Ap pSC101*ts-Rep Plac-Citrus-(+2)LAA (ssrA(+2)-tag)
pPM9 Ap pSC101*ts-Rep Ptet-weakRBS-Citrus-LAA (ssrA-tag)
pPM10 Ap pSC101*ts-Rep Plac-weakRBS-Citrus-LAA (ssrA-tag)
pPM11 Ap pSC101*ts-Rep Ptet-weakRBS-Citrus
pPM12 Ap pSC101*ts-Rep Plac-weakRBS-Citrus
pPM13 Ap pSC101 Ptet-Citrus-LAA (ssrA-tag)
pPM14 Ap pSC101 Plac-Citrus-LAA (ssrA-tag)
pPM15 Ap pSC101 Ptet-Citrus
pPM16 Ap pSC101 Plac-Citrus
pPM17 Ap pSC101 Ptet-silVenus-LAA
pPM18 Ap pSC101 Plac-silVenus-LAA
pPM19 Ap pSC101 Ptet-silVenus
pPM20 Ap pSC101 Plac-silVenus
pPM22 Ap pSC101*ts-Rep 240 lacO repeats
pPM23 Ap pSC101*ts-Rep 24 lacO repeats
pPM24 Ap pSC101*ts-Rep 24 tetO repeats
pPM25 Ap pSC101*ts-Rep 120 lacO repeats
pPM26 Ap pSC101*ts-Rep 120 tetO repeats
pPM27 LTR-5(p15A_F) Cm, Ap p15A lacI-eCFP_tetR-eYFP (bicistronic)
pPM28 LTR-13(p15A_R) Cm, Ap p15A lacI-eCFP_tetR-eYFP (bicistronic)
pPM29 Ap pSC101 Ptet-mCherry
pPM30 Ap pSC101 Plac-mCherry
pPM31 Cm, Ap p15A lacI-eYFP (removed XhoI fragment from pPM27)
pPM32 Cm p15A lacI-eCFP_tetR-eYFP (removed ApR from pPM27)
pPM33 Cm p15A lacI-eYFP (removed ApR from pPM31)
pPM34 Ap, Kn pUC19 dhl19 + Ptet-Citrus from pPM15
pPM35 Ap, Kn pUC19 dhl19 + Ptet-mCherry from pPM29
pPM36 Ap, Kn pUC19 dhl19 + Plac-Citrus from pPM16
pPM37 Ap, Kn pUC19 dhl19 + Plac-mCherry from pPM30
pPM38 Ap pSC101*ts-Rep Ptet-mCherry
pPM39 Ap pSC101*ts-Rep Plac-mCherry
pPM40 Cm F (ori2, pBeloBAC11) Ptet-Citrus
pPM41 Cm F (ori2, pBeloBAC11) Plac-Citrus
pPM42 Cm F (ori2, pBeloBAC11) Ptet-mCherry
pPM43 Ap pSC101*ts-Rep Ptet-YPet
pPM44 Ap pSC101*ts-Rep Plac-YPet
pPM45 Ap pSC101 Ptet-YPet
pPM46 Ap pSC101 Plac-YPet
pPM47 Ap pSC101*ts-Rep (pGRG36) Ptet-mCherry for integration at attTn7
pPM48 Ap pSC101*ts-Rep (pGRG36) Plac-mCherry for integration at attTn7
pPM49 forthcoming Ap R1 parA- parD+ (BLA crick)
pPM50 Ap R1 parA- parD+ (BLA watson)
pPM51 Ap R1 parA- parD- (BLA crick)
pPM52 Ap R1 parA- parD- (BLA watson)
pPM53 Kn pSC101 Plac-copA derived from pKG339
pPM54 Kn pSC101 Plac-copA derived from pKG339 (smaller than pPM53)
pPM55 Ap pSC101*ts-Rep promoter-less Citrus derived from pPM6
pPM56 Ap pSC101*ts-Rep promoter-less Dendra2 derived from pPM55, clone 56-1
pPM57 Ap pSC101*ts-Rep PLtet-Dendra derived from pPM3
pPM58 Ap pSC101*ts-Rep PLlac-Dendra derived from pPM4
pPM59 pPM51(noSTAR) Ap R1 parA- parD- (BLA crick), clone black-D10
pPM60 Ap pSC101*ts-Rep PrnaI - Citrus; -35(TTGAAG), -10(TACACT); bright
pPM61 Ap pSC101*ts-Rep PrnaI*br-1 - Citrus; -35(TTGAAG), -10(TACACT); bright
pPM62 Ap pSC101*ts-Rep PrnaI*sb-1 - Citrus; -35(TTGtAa), -10(TACACT); super bright
pPM63 Ap pSC101*ts-Rep PrnaI*sb-2 - Citrus; -35(TTGcAt), -10(TACACT); super bright
pPM64 Ap pSC101*ts-Rep PrnaI*int-1 - Citrus; -35(TgGtAa), -10(TAgACT); intermediate
pPM65 Ap pSC101*ts-Rep PrnaI*int-2 - Citrus; -35(TaGtAa), -10(TACACT); intermediate
pPM66 Ap pSC101*ts-Rep PrnaI*int-4 - Citrus; -35(TTGtAG), -10(TAgAgT); intermediate
pPM67 Ap pSC101*ts-Rep PrnaI*int-5 - Citrus; -35(TTGcAc), -10(TAgACT); intermediate
pPM68 Ap pSC101*ts-Rep PrnaI*int-7 - Citrus; -35(TTGtAG), -10(gAtACT); intermediate
pPM70 Ap pSC101*ts-Rep PrnaI - Dendra2; -35(TTGAAG), -10(TACACT); bright
pPM71 Ap pSC101*ts-Rep PrnaI*4 - Dendra2; -35(TTGAAt), -10(TACAgT); bright
pPM72 Ap pSC101*ts-Rep PrnaI*5 - Dendra2; -35(TTGAAT), -10(aAgACT); bright
pPM73 Ap pSC101*ts-Rep PrnaI*6b - Dendra2; -35(TgGAAT), -10(cAtAaTAGAAGa); dim
pPM74 Ap pSC101*ts-Rep PrnaI*7 - Dendra2; -35(TaGtAa), -10(TACACT); intermediate
pPM75 Kn pSC101(E93K) PA1/O4-copA-rrnB-LacIq from pKG339 3x higher copy number than pSC101 wt
pPM76 Kn pSC101 PA1/O4-copA-rrnB-LacIq from pKG339 backbone is Uri Alon's pUA66

Rack II

Name Alias Marker(s) Origin Insert Notes
pPM77 Ap pSC101 add LacIq to pPM30 (LacIq PCR prod. from pKG339 (seq. NOT fully checked, but functional))
pPM78 Ap pSC101 add LacIq to pPM16 (LacIq PCR prod. from pKG339 (seq. NOT fully checked, but functional))
pPM79 491ΔR Ap R1 (ts-runaway) par+ pKG491 ΔEcoRI site
pPM80-2 491ΔR-B Ap R1 (ts-runaway) par- pKG491 Δ(EcoRI-BamHI+131bp)(EcoRI site recreated)
pPM80-3 491ΔR-B Ap R1 (ts-runaway) par- pKG491 Δ(~3.9kb+EcoRI-BamHI)
pPM80-5 491ΔR-B Ap R1 (ts-runaway) par- pKG491 Δ(~2.9kb+EcoRI-BamHI)
pPM80-6 491ΔR-B Ap R1 (ts-runaway) par- pKG491 Δ(~1.9kb+EcoRI-BamHI)
pPM81 491ΔRΔB Ap R1 (ts-runaway) par+ pPM79 ΔBamHI site
pPM82 54ΔAcu Kan pSC101 pPM54 ΔAcuI, removes duplicated sequences present in pPM54
pPM83 75parΔBsp Kn pSC101(E93K) PA1/O4-copA-rrnB-LacIq from pKG339 removes duplicated sequences from pPM75, 3x higher copy number than pSC101 wt
pPM84 15-PA1/O4 Ap pSC101 PA1/O4-Citrus very strong promoter, makes cells sick
pPM85 29-PA1/O4 Ap pSC101 PA1/O4-mCherry very strong promoter, makes cells sick (but less than pPM84)
pPM86 84+lac Ap pSC101 PA1/O4-Citrus; LacIq
pPM87 85+lac Ap pSC101 PA1/O4-mCherry; LacIq
pPM88 78.PA1/O4 Ap pSC101 PA1/O4-Citrus; LacIq
pPM89 77.PA1/O4 Ap pSC101 PA1/O4-mCherry; LacIq
pPM90 491ΔR-B Ap R1 (ts-runaway) par- clone 4; pPM80-2 with EcoRI site deleted
pPM91 491ΔRΔBΔAfe Ap R1 (ts-runaway) par+ pPM81 cut with AfeI, Afe-XhoI-SacI-AfeI cassette inserted
pPM91-3 (flipped) 491ΔRΔBΔAfe Ap R1 (ts-runaway) par+ pPM81 cut with AfeI, Afe-XhoI-SacI-AfeI cassette inserted, cassette orientation flipped relative to pPM91
pPM92 (flipped) 491ΔR-BΔAfe Ap R1 (ts-runaway) par- pPM90 cut with AfeI, Afe-XhoI-SacI-AfeI cassette inserted, cassette orientation flipped relative to pPM91
pPM93 81+15 Ap R1 (ts-runaway) par+ pPM81 with PLtet-Citrus from pPM15
pPM94 81+70 Ap R1 (ts-runaway) par+ pPM81 with PRNAI-Dendra2 from pPM70
pPM95 90+15 Ap R1 (ts-runaway) par- pPM90 with PLtet-Citrus from pPM15
pPM96 90+70 Ap R1 (ts-runaway) par- pPM90 with PRNAI-Dendra2 from pPM70
pPM97 R1 from 141 Ap R1 (ts-runaway) par+ pDNA purified from PMB141, transformed into MC1061, select for AmpR, grow and purify plasmid
pPM98 R1 from 197 Ap R1 (ts-runaway) par+ pDNA purified from PMB197, transformed into MC1061, select for AmpR, grow and purify plasmid
pPM99 91+15 Ap R1 (ts-runaway) par+ pPM91 with PLtet-Citrus insert from pPM15
pPM100 91+70 Ap R1 (ts-runaway) par+ pPM91 with PRNAI-Dendra2 insert from pPM70
pPM101 92+15 Ap R1 (ts-runaway) par- pPM92 with PLtet-Citrus insert from pPM15
pPM102 92+70 Ap R1 (ts-runaway) par- pPM92 with PRNAI-Dendra2 insert from pPM70
pPM103A 12+86 Kn pSC101(E93K) PA1/O4-Citrus-rrnB-LacIq from pPM86 in pJPK12 103A and B differ in orientation of insert
pPM103B 12+86 Kn pSC101(E93K) PA1/O4-Citrus-rrnB-LacIq from pPM86 in pJPK12 103A and B differ in orientation of insert
pPM104
pPM105
pPM106A 54(Sal)+75 Kn pSC101 2x(PA1/O4-copA-rrnB term-LacIq) A and B differ in orientation of insert
pPM107A 54(Sal)+83 Kn pSC101 2x(PA1/O4-copA-rrnB term-LacIq) A and B differ in orientation of insert
pPM108
pPM109
pPM110A 82(Sca)+ST95 Kn pSC101 1x(PA1/O4-copA-rrnB term-LacIq) + 1x(PLtet-copA-T1 term-TetR) A and B differ in orientation of insert
pPM110B 82(Sca)+ST95 Kn pSC101 1x(PA1/O4-copA-rrnB term-LacIq) + 1x(PLtet-copA-T1 term-TetR) A and B differ in orientation of insert
pPM111A 54(Sca)+ST95 Kn pSC101 1x(PA1/O4-copA-rrnB term-LacIq) + 1x(PLtet-copA-T1 term-TetR) A and B differ in orientation of insert
pPM112 93 w/ PL Ap R1 (ts-runaway) par+ Plambda-Citrus; made by promoter replacement of pPM93
pPM113 95 w/ PL Ap R1 (ts-runaway) par- Plambda-Citrus; made by promoter replacement of pPM95
pPM114 112 del AgeI Ap R1 (ts-runaway) par+ clone 1
pPM115 113 del AgeI Ap R1 (ts-runaway) par-
pPM116 Ap R1 (ts-runaway?) par- 32G mutation introduced in PL upstream of copB-delK-mutant
pPM117 Ap R1 (ts-runaway?) par- 41C mutation introduced in PL upstream of copB-delK-mutant
pPM118 Ap R1 (ts-runaway?) par- 32G & 41C mutations introduced in PL upstream of copB-delK-mutant
pPM119 Ap R1 (ts-runaway?) par- 32G mutation introduced in PL upstream of copB-delK-mutant
pPM120 Ap R1 (ts-runaway?) par- 41C mutation introduced in PL upstream of copB-delK-mutant
pPM121* Ap R1 (ts-runaway?) par- 32G & 41C mutations introduced in PL upstream of copB-delK-mutant (*also single bp del in primer)
pPM122 113+32G Ap R1 (ts-runaway?) par- 32G mutation introduced in PL upstream of copB
pPM123 113+41C Ap R1 (ts-runaway?) par- 41C mutation introduced in PL upstream of copB
pPM124 113+32G/41C Ap R1 (ts-runaway?) par- 32G & 41C mutations introduced in PL upstream of copB
pPM125 90+32G Ap R1 (ts-runaway?) par- 32G mutation introduced in PL upstream of copB
pPM126 90+41C Ap R1 (ts-runaway?) par- 41C mutation introduced in PL upstream of copB
pPM127 90+32G/41C Ap R1 (ts-runaway?) par- 32G & 41C mutations introduced in PL upstream of copB
pPM128 81+Hpa Ap R1 par+ PL-cI857 replaced with AflII-HpaI fragment from pKN1562 (this insert contains some seq. from R1 & some from Tn802)
pPM129 81+Afe Ap R1 par+ PL-cI857 replaced with AflII-AfeI fragment from pKN1562 (this insert contains some seq. from R1 & some from Tn802)
pPM130 112+Psh Ap R1 par+ PL-cI857 replaced with AflII-PshAI fragment from pKN1562 (this insert contains some seq. from R1 & some from Tn802)
pPM131 112+Hpa Ap R1 par+ PL-cI857 replaced with AflII-HpaI fragment from pKN1562 (this insert contains some seq. from R1 & some from Tn802)
pPM132 112+Afe Ap R1 par+ PL-cI857 replaced with AflII-AfeI fragment from pKN1562 (this insert contains some seq. from R1 & some from Tn802)
pPM133 113+Hpa Ap R1 par- PL-cI857 replaced with AflII-HpaI fragment from pKN1562 (this insert contains some seq. from R1 & some from Tn802)
pPM134 113+Afe Ap R1 par- PL-cI857 replaced with AflII-AfeI fragment from pKN1562 (this insert contains some seq. from R1 & some from Tn802)
pPM135 132+X_R Ap R1 par+ PL promoter replaced w/ promoter-less oligo cassette
pPM136 132+X_T7_R Ap R1 par+ PL promoter replaced w/ T7 promoter cassette
pPM137 132+PL(wt) Ap R1 par+ PL promoter replaced w/ cI857-PL(wt)
pPM138 132+PL(32G) Ap R1 par+ PL promoter replaced w/ cI857-PL*(32G)
pPM139 132+PL(41C) Ap R1 par+ PL promoter replaced w/ cI857-PL*(41C)
pPM140 134+X_T7_R Ap R1 par- PL promoter replaced w/ T7 promoter cassette
pPM141 134+PL(41C) Ap R1 par- PL promoter replaced w/ cI857-PL*(41C)
pPM142 134+PL(wt) Ap R1 par- PL promoter replaced w/ cI857-PL(wt)

Rack III

Name Alias Marker(s) Origin Insert Notes
pPM144 Ap R1 Pt7-Citrus clone-3; isothermally assembled (Aa-), all PCR; fully sequenced
pPM145 Ap R1 Pt7-Citrus; parMRC clone-A+5; isothermally assembled (Aa+), all PCR; restruck on LB/Ap20 to avoid selection of copy number mutants; fully sequenced
pPM146 Ap R1 Pλ-Citrus clone-1; isothermally assembled (Aλa-), all PCR
pPM147 Ap R1 Pλ-Citrus; parMRC clone-2; isothermally assembled (Aλa+), all PCR; BEWARE copy number mutants
pPM148 Ap R1 Pλ-Citrus clone-1; isothermally assembled (Ba-), ori and FP from pPM130
pPM149 Ap R1 Pλ-Citrus; parMRC clone-1; isothermally assembled (Ba+), ori and FP from pPM130
pPM150 Cm R1 Pt7-Citrus clone-1; isothermally assembled (Ac-), all PCR; transformants selected on 15ug/ml Cm
pPM151 Cm R1 Pλ-Citrus clone-1; isothermally assembled (Aλc-), all PCR; transformants selected on 15ug/ml Cm
pPM152 Ap R1 Pt7-mCherry pPM144 with mCherry from DHL374 (BamHI-XbaI)
pPM153 Ap R1 Pt7-GFP pPM144 with gfpmut3 from DHL408 (BamHI-XbaI)
pPM154 Ap R1 Pt7-mCherry; parMRC pPM145 with mCherry from DHL374 (BamHI-XbaI); BEWARE copy number mutants
pPM155 Ap R1 Pt7-GFP; parMRC pPM145 with gfpmut3 from DHL408 (BamHI-XbaI); BEWARE copy number mutants
pPM156 Cm R1 Pt7-Citrus clone-1; isothermally assembled (lAc-), all PCR; transformants selected on 3ug/ml Cm
pPM158 Cm R1 Pt7-Citrus clone-1; isothermally assembled (lBc-), ori and FP from pPM130; transformants selected on 3ug/ml Cm
pPM160 Cm R1 Pt7-mCherry clone-1; pPM156 with mCherry from DHL374 (BamHI-XbaI)
pPM161 Cm R1 Pt7-GFP clone-1; pPM156 with gfpmut3 from DHL408 (BamHI-XbaI)
pPM162 Ap R1 Pt7-Citrus clone-1; pPM148 with Pλ replaced by Pt7
pPM163 Ap R1 Pt7-Citrus; parMRC clone-1; pPM149 with Pλ replaced by Pt7
pPM164 Ap R1 Pt7-Citrus; parC ΔparMR pPM145-A5 Δbp#3450-3990; digest gave unexpected pattern
pPM165 Ap R1 Pt7-Citrus; parCM ΔparR clone-4; pPM145-A5 ΔHpaI-BsaBI (bp#3622-3694)
pPM166 Ap R1 Pt7-Citrus; parMRC clone 25-6; pPM145-A5 BLA flipped (BspHI)
pPM167 Ap R1 Pt7-Citrus; parMRC clone 80-6; pPM145-A5 2xBLA (BspHI)
pPM168 Ap R1 Pt7-Citrus; parMRC clone 80-7; pPM145-A5 2xBLA flipped (BspHI)
pPM169 Ap R1 Pt7-Citrus; parMRC clone T1-1; pPM145 with 167bp deletion of ycdAB
pPM170 Ap R1 Pt7-Citrus; parMRC clone T2-1; pPM145 with 116bp deletion of ycdAB
pPM171 Ap R1 Pt7-Citrus; parMRC clone T3-1; pPM145 with 344bp deletion of ycdAB
pPM172 Ap R1 Pt7-Citrus; parMRC clone E1-1; pPM145 with 600bp S.pombe DNA btwn. bla and par
pPM173 Ap R1 Pt7-Citrus; parMRC clone E2-1; pPM145 with 500bp S.pombe DNA btwn. bla and par
pPM174 Ap R1 Pt7-Citrus; parMRC clone E3-1; pPM145 with 1kb S.pombe DNA btwn. bla and par
pPM175 Ap R1 Pt7-Citrus; parMRC pPM170 with 500bp S.pombe DNA btwn. bla and par
pPM176 Ap R1 Pt7-Citrus; parMRC pPM170 with 1kb S.pombe DNA btwn. bla and par
pPM177 Ap R1 Pt7-Citrus; parMRC pPM171 with 500bp S.pombe DNA btwn. bla and par
pPM178 Ap R1 Pt7-Citrus; parMRC pPM171 with 1kb S.pombe DNA btwn. bla and par
pPM179 Ap R1 Pt7-Citrus; parCM ΔparR pPM170 ΔHpaI-BsaBI (bp#3622-3694)
pPM180 Ap R1 Pt7-RFP; parMRC pPM170, replaced Citrus with Dirk's mCherry
pPM181 Ap R1 Pt7-GFP; parMRC pPM170, replaced Citrus with gfpmut3
pPM182 Spc pSC101 PA1/O4-copA from pNG162 (attenuated Ptrc); PA1/O4-copA insert from pKG339 results in duplicated seq. with pNG162 bkbone
pPM183 Spc pSC101 PA1/O4-copA from pNG162a (full-strength Ptrc); PA1/O4-copA insert from pKG339 results in duplicated seq. with pNG162 bkbone
pPM184 Spc pSC101 Ptrc-copB from pNG162 (attenuated Ptrc); copB extends into copT, binding site for copA
pPM185 Spc pSC101 Ptrc-copB from pNG162a (full-strength Ptrc); copB extends into copT hairpin, binding site for copA
pPM186 Spc pSC101 Ptrc-copB, PA1/O4-copA from pNG162 (attenuated Ptrc); duplication and copT removed, but no trx terminator between copB and copA; tested 110818: arrests better than pPM182, but worse than pPM54
pPM187 Spc pSC101 Ptrc-copB, PA1/O4-copA from pNG162a (full-strength Ptrc); duplication and copT removed, but no trx terminator between copB and copA; tested 110818: arrests better than pPM182, but worse than pPM186
pPM188 Spc pSC101 Ptrc-copB from pNG162 (attenuated Ptrc); removed copT from pPM184
pPM189 Spc pSC101 Ptrc-copB from pNG162a (full-strength Ptrc); removed copT from pPM185
pPM190 Kn pSC101 PA1/04-copA + P204-copB pPM54 + attenuated Ptrc-copB-rrnB from pPM188
pPM191 Kn pSC101 PA1/04-copA + P99a-copB pPM54 + Ptrc-copB-rrnB from pPM189
pPM192 Kn pSC101 2x(PA1/04-copA_lacIq) + P204-copB pPM107A + attenuated Ptrc-copB-rrnB from pPM188
pPM193 Kn pSC101 2x(PA1/04-copA_lacIq) + P99a-copB pPM107A + Ptrc-copB-rrnB from pPM189
pPM194 parMstart-RFP-parR Ap R1 mCherry-mKate2 hybrid in place of parM clone A1; made by isothermal assembly; partially sequenced
pPM195 parMstart-RFP-deltaR Ap R1 mCherry-mKate2 hybrid in place of parM; ΔparR clone B2; made by isothermal assembly; partially sequenced
pPM196 parMstart-stop_startRFP-parR Ap R1 mCherry-mKate2 hybrid after parM 1-30aa clone C1; made by isothermal assembly; partially sequenced
pPM197 parMstart-stop_startRFP-deltaR Ap R1 mCherry-mKate2 hybrid after parM 1-30aa; ΔparR clone D2; made by isothermal assembly; partially sequenced
pPM198 pKGCm2x Cm p15A (2x)PA1/O4-copA_rrnB_lacIq II+ clone 1;
pPM199 54Spc Spc pSC101 PA1/O4-copA_rrnB_lacIq clone 1; replaced KanR in pPM54 with SpcR
pPM200 106Spc Spc pSC101 (2x)PA1/O4-copA_rrnB_lacIq clone 1; replaced KanR in pPM106A with SpcR
pPM201 107Spc Spc pSC101 (2x)PA1/O4-copA_rrnB_lacIq clone 1; replaced KanR in pPM107A with SpcR
pPM202 153Kn Kn R1 Pt7-GFP (gfpmut3) clone 1; pPM153 replaced AmpR with KanR
pPM203 181Kn Kn R1 Pt7-GFP (gfpmut3); parMRC clone 1; pPM181 replaced AmpR with KanR
pPM204 144K2 Ap R1 Pt7-mKate2* clone 1; pPM144 replaced Citrus with *mCherry-mKate2 hybrid from pNDL-194
pPM205 170K2 Ap R1 Pt7-mKate2*; parMRC clone 1; pPM170 replaced Citrus with *mCherry-mKate2 hybrid from pNDL-194
pPM208 mV2 Ap R1 parC__mCherry-mKate2 hybrid in place of parM__parR-mVenus (C-term fusion) not made
pPM209 mV3 Ap R1 parC__mCherry-mKate2 hybrid in place of parM__parR-mVenus (Δ20aa C-term fusion) clone V3-1; made by isothermal assembly; partially sequenced
pPM210 parMstart-YFP-parR Ap R1 Citrus in place of parM clone 1-2; made by isothermal assembly; partially sequenced
pPM211 parR-mV5 Ap R1 parC__mCherry-mKate2__parR(1-56)-mVenus clone 2-3; made by isothermal assembly; partially sequenced
pPM212 parR-mV4 Ap R1 parC__mCherry-mKate2__parR(1-69)-mVenus clone 3-2; made by isothermal assembly; partially sequenced
pPM213 parR-Venus Ap R1 parC__mCherry-mKate2__parR(1-97)-Venus clone 4-1; made by isothermal assembly; partially sequenced
pPM214 parR-sfGFP Ap R1 parC__mCherry-mKate2__parR(1-97)-sfGFP clone 5-3; made by isothermal assembly; partially sequenced
pPM215 parR-mEGFP Ap R1 parC__mCherry-mKate2__parR(1-97)-mEGFP clone 6-1; made by isothermal assembly; partially sequenced
pPM216 G3 Ap R1 parC__mCherry-mKate2 hybrid in place of parM__parR(1-97)-mGFPmut3 clone 8-1; made by isothermal assembly; partially sequenced
pPM217 Ap pSC101-ts parMstart_RFP_parR from pPM194 inserted in pGRG36 for integration at attTn7
pPM218 Ap pSC101-ts parMstart_RFP_deltaR from pPM195 inserted in pGRG36 for integration at attTn7
pPM219 Ap pSC101-ts parMstart_RFP_parR(1-97)-mVenus from pPM209 inserted in pGRG36 for integration at attTn7
pPM220 Ap pSC101-ts parMstart_RFP_parR(1-97)-Venus from pPM213 inserted in pGRG36 for integration at attTn7
pPM221 Ap pSC101-ts parMstart_RFP_parR(1-97)-sfGFP from pPM214 inserted in pGRG36 for integration at attTn7
pPM222 Ap R1 pPM136 + lacIq-ssrA for measuring plasmid loss with Billy's assay
pPM223 Ap R1 pPM140 + lacIq-ssrA for measuring plasmid loss with Billy's assay

Rack IV

Name Alias Marker(s) Origin Insert Notes
pPM224 pDAG203+194 Cm oriF(2) parC_RFP_parR
pPM225 pDAG203+195 Cm oriF(2) parC_RFP
pPM226 pDAG203+213 Cm oriF(2) parC_RFP_parR(1-97)-Venus
pPM227 pDAG203+214 Cm oriF(2) parC_RFP_parR(1-97)-sfGFP
pPM228 158_cop12_dT7 Cm oriR1_copA-12 mutation point mutation in copA (~4x copy-up, compatible with wt copA); Pt7 promoter removed

T-series plasmids

Generated by digesting backbone (# before T) with ClaI and non-directional insertion of ClaI digested TetR PCR product. Selected non-fluorescent transformants. Apparently ClaI had STAR activity and religation yielded many plasmids that were non-fluorescent because they had lost the YFP sequence, not because they had TetR. TetR PCR product has not been sequenced.

Name Alias Marker(s) Origin Insert Notes
pPM93T-1 Ap R1 (ts runaway) par+ ClaI to just upstream of cI857 deleted
pPM93T-2 Ap R1 (ts runaway) par+; TetR inserted at ClaI, coding strand same as repA
pPM95T-2 Ap R1 (ts runaway) par- ClaI to just upstream of copB deleted
pPM95T-3 Ap R1 (ts runaway) par- ClaI to just upstream of cI857 deleted
pPM95T-7 Ap R1 (ts runaway) par-; TetR inserted at ClaI, coding strand opposite repA
pPM95T-8 Ap R1 (ts runaway) par-; TetR inserted at ClaI, coding strand same as repA
pPM99T-1 Ap R1 (ts runaway) par+; TetR inserted at ClaI, coding strand same as repA
pPM99T-2 Ap R1 (ts runaway) par+; TetR inserted at ClaI, coding opposite repA
pPM101T-1 Ap R1 (ts runaway) par- ClaI to just upstream of copB deleted
pPM101T-2 Ap R1 (ts runaway) par-; TetR inserted at ClaI, coding same as repA
pPM101T-3 Ap R1 (ts runaway) par- ClaI to halfway into copB deleted


@#&^%^$'s plasmids

Name Alias Origin Insert
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