pEXP3-DEST
Gateway® destination vector for cell-free expression and N-terminal tagging. See also pEXP1-DEST.
Sequence Author: Thermo Fisher (Invitrogen)
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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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Sticky ends from different AflIII sites may not be compatible. |
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PciI is inhibited by nonionic detergents. |
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After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility. |
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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. |
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The 1-base overhangs produced by BmrI may be hard to ligate.Sticky ends from different BmrI sites may not be compatible.Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium. |
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Efficient cleavage requires at least two copies of the NmeAIII recognition sequence. Sticky ends from different NmeAIII sites may not be compatible.For full activity, add fresh S-adenosylmethionine (SAM). |
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BsaHI is typically used at 37°C, but is even more active at 60°C. |
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Sticky ends from different DraIII sites may not be compatible. |
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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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The 1-base overhangs produced by XcmI may be hard to ligate.Sticky ends from different XcmI sites may not be compatible. |
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Sticky ends from different BsmBI sites may not be compatible.BsmBI-v2 is an improved version of BsmBI. |
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Sticky ends from different Esp3I sites may not be compatible. |
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Sticky ends from different PasI sites may not be compatible. |
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| * Blocked by Dcm methylation. Sticky ends from different PflMI sites may not be compatible. |
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BssHII is typically used at 50°C, but is 75% active at 37°C. |
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Sticky ends from different AvaI sites may not be compatible. |
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Sticky ends from different BsoBI sites may not be compatible.BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C. |
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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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Efficient cleavage requires at least two copies of the BfuAI recognition sequence. Sticky ends from different BfuAI sites may not be compatible.BfuAI is typically used at 50°C, but is 50% active at 37°C. |
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Efficient cleavage requires at least two copies of the BspMI recognition sequence. Sticky ends from different BspMI sites may not be compatible. |
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Sticky ends from different BlpI sites may not be compatible. |
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Sticky ends from different EcoO109I sites may not be compatible. |
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Efficient cleavage requires at least two copies of the NgoMIV recognition sequence. |
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Efficient cleavage requires at least two copies of the NaeI recognition sequence. |
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Sticky ends from different BanII sites may not be compatible. |
| AmpR 2753 .. 3613 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 2753 .. 2821 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 2753 .. 3613 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 2822 .. 3613 = 792 bp 263 amino acids = 29.0 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 2753 .. 3613 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| CmR 420 .. 1079 = 660 bp 219 amino acids = 25.7 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol |
| CmR 420 .. 1079 = 660 bp 219 amino acids = 25.7 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol |
| ori 3784 .. 4372 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 3784 .. 4372 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| f1 ori 2112 .. 2567 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| f1 ori 2112 .. 2567 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| ccdB 1421 .. 1726 = 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 1421 .. 1726 = 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. |
| attR1 187 .. 311 = 125 bp recombination site for the Gateway® LR reaction |
| attR1 187 .. 311 = 125 bp recombination site for the Gateway® LR reaction |
| attR2 1767 .. 1891 = 125 bp recombination site for the Gateway® LR reaction |
| attR2 1767 .. 1891 = 125 bp recombination site for the Gateway® LR reaction |
| AmpR promoter 2648 .. 2752 = 105 bp |
| AmpR promoter 2648 .. 2752 = 105 bp |
| ATG 100 .. 102 = 3 bp 1 amino acid = 149.2 Da |
| ATG 100 .. 102 = 3 bp 1 amino acid = 149.2 Da |
| 6xHis 103 .. 120 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 103 .. 120 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| tetracysteine tag 133 .. 150 = 18 bp 6 amino acids = 584.8 Da Product: tetracysteine peptide that binds biarsenical labeling reagents |
| tetracysteine tag 133 .. 150 = 18 bp 6 amino acids = 584.8 Da Product: tetracysteine peptide that binds biarsenical labeling reagents |
| TEV site 160 .. 180 = 21 bp 7 amino acids = 869.9 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| TEV site 160 .. 180 = 21 bp 7 amino acids = 869.9 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| T7 terminator 1968 .. 2015 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| T7 terminator 1968 .. 2015 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| lac UV5 promoter 336 .. 366 = 31 bp 3 segments Segment 1: -35 336 .. 341 = 6 bp E. coli lac promoter with an "up" mutation |
| lac UV5 promoter 336 .. 366 = 31 bp 3 segments Segment 2: 342 .. 359 = 18 bp E. coli lac promoter with an "up" mutation |
| lac UV5 promoter 336 .. 366 = 31 bp 3 segments Segment 3: -10 360 .. 366 = 7 bp E. coli lac promoter with an "up" mutation |
| lac UV5 promoter 336 .. 366 = 31 bp 3 segments E. coli lac promoter with an "up" mutation |
| RBS 70 .. 92 = 23 bp efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989) |
| RBS 70 .. 92 = 23 bp efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989) |
| T7 promoter 20 .. 38 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 20 .. 38 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| ORF: 1421 .. 1726 = 306 bp ORF: 101 amino acids = 11.7 kDa |
| ORF: 2753 .. 3613 = 861 bp ORF: 286 amino acids = 31.6 kDa |
| ORF: 420 .. 1079 = 660 bp ORF: 219 amino acids = 25.7 kDa |
| ORF: 3217 .. 3483 = 267 bp ORF: 88 amino acids = 9.2 kDa |
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