Paulsson:Plasmids
From OpenWetWare
Jump to navigationJump to search
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 |
|---|
