pET-DEST42
Gateway® destination vector for high-level T7-regulated bacterial expression and C-terminal tagging.
Sequence Author: Thermo Fisher (Invitrogen)
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The 1-base overhangs produced by EcoNI may be hard to ligate.Sticky ends from different EcoNI sites may not be compatible. |
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Sticky ends from different BstEII sites may not be compatible.BstEII is typically used at 60°C, but is 50% active at 37°C. |
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ApaI can be used between 25°C and 37°C. |
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PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations. |
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The 1-base overhangs produced by PflFI may be hard to ligate.Sticky ends from different PflFI sites may not be compatible. |
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The 1-base overhangs produced by Tth111I may be hard to ligate.Sticky ends from different Tth111I sites may not be compatible. |
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Sticky ends from different BspQI sites may not be compatible. |
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Sticky ends from different SapI sites may not be compatible.SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot. |
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PciI is inhibited by nonionic detergents. |
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Efficient cleavage requires at least two copies of the SgrAI recognition sequence. |
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Sticky ends from different PasI sites may not be compatible. |
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Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. |
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SmaI can be used at 37°C for brief incubations. |
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This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site. |
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Sticky ends from different BseRI sites may not be compatible.BseRI quickly loses activity at 37°C.Prolonged incubation with BseRI may lead to degradation of the DNA. |
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Sticky ends from different BlpI sites may not be compatible. |
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The 1-base overhangs produced by AhdI may be hard to ligate. Sticky ends from different AhdI sites may not be compatible. |
lacI 6289 .. 7371 = 1083 bp 360 amino acids = 38.6 kDa Product: lac repressor The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG). |
lacI 6289 .. 7371 = 1083 bp 360 amino acids = 38.6 kDa Product: lac repressor The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG). |
AmpR 2737 .. 3597 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 1: signal sequence 2737 .. 2805 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
AmpR 2737 .. 3597 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 2: 2806 .. 3597 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
AmpR 2737 .. 3597 = 861 bp 286 amino acids = 31.5 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
CmR 635 .. 1294 = 660 bp 219 amino acids = 25.7 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol |
CmR 635 .. 1294 = 660 bp 219 amino acids = 25.7 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol |
ori 3768 .. 4356 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
ori 3768 .. 4356 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
ccdB 1636 .. 1941 = 306 bp 101 amino acids = 11.7 kDa Product: CcdB, a bacterial toxin that poisons DNA gyrase Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains. |
ccdB 1636 .. 1941 = 306 bp 101 amino acids = 11.7 kDa Product: CcdB, a bacterial toxin that poisons DNA gyrase Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains. |
rop 4786 .. 4977 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
rop 4786 .. 4977 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
attR1 402 .. 526 = 125 bp recombination site for the Gateway® LR reaction |
attR1 402 .. 526 = 125 bp recombination site for the Gateway® LR reaction |
attR2 1982 .. 2106 = 125 bp recombination site for the Gateway® LR reaction |
attR2 1982 .. 2106 = 125 bp recombination site for the Gateway® LR reaction |
AmpR promoter 2632 .. 2736 = 105 bp |
AmpR promoter 2632 .. 2736 = 105 bp |
lacI promoter 7372 .. 9 = 78 bp |
lacI promoter 7372 .. 9 = 78 bp |
V5 tag 2126 .. 2167 = 42 bp 14 amino acids = 1.4 kDa Product: epitope tag from simian virus 5 |
V5 tag 2126 .. 2167 = 42 bp 14 amino acids = 1.4 kDa Product: epitope tag from simian virus 5 |
6xHis 2177 .. 2194 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
6xHis 2177 .. 2194 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
T7 terminator 2265 .. 2312 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
T7 terminator 2265 .. 2312 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
lac UV5 promoter 551 .. 581 = 31 bp 3 segments Segment 1: -35 551 .. 556 = 6 bp E. coli lac promoter with an "up" mutation |
lac UV5 promoter 551 .. 581 = 31 bp 3 segments Segment 2: 557 .. 574 = 18 bp E. coli lac promoter with an "up" mutation |
lac UV5 promoter 551 .. 581 = 31 bp 3 segments Segment 3: -10 575 .. 581 = 7 bp E. coli lac promoter with an "up" mutation |
lac UV5 promoter 551 .. 581 = 31 bp 3 segments E. coli lac promoter with an "up" mutation |
lac operator 337 .. 361 = 25 bp The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG). |
lac operator 337 .. 361 = 25 bp The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG). |
T7 promoter 318 .. 336 = 19 bp promoter for bacteriophage T7 RNA polymerase |
T7 promoter 318 .. 336 = 19 bp promoter for bacteriophage T7 RNA polymerase |
ORF: 1636 .. 1941 = 306 bp ORF: 101 amino acids = 11.7 kDa |
ORF: 2737 .. 3597 = 861 bp ORF: 286 amino acids = 31.5 kDa |
ORF: 107 .. 346 = 240 bp ORF: 79 amino acids = 8.0 kDa |
ORF: 635 .. 1294 = 660 bp ORF: 219 amino acids = 25.7 kDa |
ORF: 5009 .. 5377 = 369 bp ORF: 122 amino acids = 14.2 kDa |
ORF: 5811 .. 6506 = 696 bp ORF: 231 amino acids = 25.4 kDa |
ORF: 7410 .. 233 = 264 bp ORF: 87 amino acids = 9.5 kDa |
ORF: 4786 .. 5010 = 225 bp ORF: 74 amino acids = 8.5 kDa |
ORF: 5374 .. 5712 = 339 bp ORF: 112 amino acids = 12.6 kDa |
ORF: 6289 .. 7248 = 960 bp ORF: 319 amino acids = 34.1 kDa |
ORF: 3201 .. 3467 = 267 bp ORF: 88 amino acids = 9.2 kDa |
ORF: 5616 .. 6233 = 618 bp ORF: 205 amino acids = 21.8 kDa |
ORF: 7121 .. 181 = 501 bp ORF: 166 amino acids = 17.5 kDa |
ORF: 6269 .. 6532 = 264 bp ORF: 87 amino acids = 8.9 kDa |
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