pET104-DEST
Gateway® destination vector for high-level inducible expression of biotinylated proteins in bacteria.
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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| * Blocked by Dam methylation. BclI is typically used at 50-55°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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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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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 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 6358 .. 7440 = 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 6358 .. 7440 = 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 2806 .. 3666 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 1: signal sequence 2806 .. 2874 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 2806 .. 3666 = 861 bp 286 amino acids = 31.5 kDa 2 segments Segment 2: 2875 .. 3666 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 2806 .. 3666 = 861 bp 286 amino acids = 31.5 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| CmR 794 .. 1453 = 660 bp 219 amino acids = 25.7 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol |
| CmR 794 .. 1453 = 660 bp 219 amino acids = 25.7 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol |
| ori 3837 .. 4425 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 3837 .. 4425 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ccdB 1774 .. 2079 = 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 1774 .. 2079 = 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. |
| ATG 297 .. 299 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| ATG 297 .. 299 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
| BioEase™ tag 300 .. 515 = 216 bp 72 amino acids = 7.1 kDa 3 segments Segment 1: 300 .. 410 = 111 bp 37 amino acids = 3.6 kDa Product: biotinylated sequence derived from the C-terminus of the α-subunit of Klebsiella pneumoniae oxaloacetate decarboxylase contains a unique biotinylation site |
| BioEase™ tag 300 .. 515 = 216 bp 72 amino acids = 7.1 kDa 3 segments Segment 2: site 411 .. 413 = 3 bp 1 amino acid = 146.2 Da Product: biotinylated sequence derived from the C-terminus of the α-subunit of Klebsiella pneumoniae oxaloacetate decarboxylase contains a unique biotinylation site |
| BioEase™ tag 300 .. 515 = 216 bp 72 amino acids = 7.1 kDa 3 segments Segment 3: 414 .. 515 = 102 bp 34 amino acids = 3.4 kDa Product: biotinylated sequence derived from the C-terminus of the α-subunit of Klebsiella pneumoniae oxaloacetate decarboxylase contains a unique biotinylation site |
| BioEase™ tag 300 .. 515 = 216 bp 72 amino acids = 7.1 kDa 3 segments Product: biotinylated sequence derived from the C-terminus of the α-subunit of Klebsiella pneumoniae oxaloacetate decarboxylase contains a unique biotinylation site |
| Xpress™ tag 528 .. 551 = 24 bp 8 amino acids = 998.0 Da 2 segments Segment 1: 528 .. 536 = 9 bp 3 amino acids = 409.4 Da Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site |
| Xpress™ tag 528 .. 551 = 24 bp 8 amino acids = 998.0 Da 2 segments Segment 2: enterokinase site 537 .. 551 = 15 bp 5 amino acids = 606.5 Da Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site |
| Xpress™ tag 528 .. 551 = 24 bp 8 amino acids = 998.0 Da 2 segments Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site |
| rop 4855 .. 5046 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
| rop 4855 .. 5046 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
| attR1 561 .. 685 = 125 bp recombination site for the Gateway® LR reaction |
| attR1 561 .. 685 = 125 bp recombination site for the Gateway® LR reaction |
| attR2 2120 .. 2244 = 125 bp recombination site for the Gateway® LR reaction |
| attR2 2120 .. 2244 = 125 bp recombination site for the Gateway® LR reaction |
| AmpR promoter 2701 .. 2805 = 105 bp |
| AmpR promoter 2701 .. 2805 = 105 bp |
| lacI promoter 7441 .. 7518 = 78 bp |
| lacI promoter 7441 .. 7518 = 78 bp |
| T7 terminator 2330 .. 2377 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| T7 terminator 2330 .. 2377 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| lac UV5 promoter 710 .. 740 = 31 bp 3 segments Segment 1: -35 710 .. 715 = 6 bp E. coli lac promoter with an "up" mutation |
| lac UV5 promoter 710 .. 740 = 31 bp 3 segments Segment 2: 716 .. 733 = 18 bp E. coli lac promoter with an "up" mutation |
| lac UV5 promoter 710 .. 740 = 31 bp 3 segments Segment 3: -10 734 .. 740 = 7 bp E. coli lac promoter with an "up" mutation |
| lac UV5 promoter 710 .. 740 = 31 bp 3 segments E. coli lac promoter with an "up" mutation |
| lac operator 228 .. 252 = 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 228 .. 252 = 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 209 .. 227 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 209 .. 227 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| RBS 282 .. 288 = 7 bp ribosome binding site |
| RBS 282 .. 288 = 7 bp ribosome binding site |
| ORF: 1774 .. 2079 = 306 bp ORF: 101 amino acids = 11.7 kDa |
| ORF: 2806 .. 3666 = 861 bp ORF: 286 amino acids = 31.5 kDa |
| ORF: 794 .. 1453 = 660 bp ORF: 219 amino acids = 25.7 kDa |
| ORF: 5078 .. 5446 = 369 bp ORF: 122 amino acids = 14.2 kDa |
| ORF: 7616 .. 237 = 240 bp ORF: 79 amino acids = 8.0 kDa |
| ORF: 297 .. 596 = 300 bp ORF: 99 amino acids = 10.1 kDa |
| ORF: 5880 .. 6575 = 696 bp ORF: 231 amino acids = 25.4 kDa |
| ORF: 7479 .. 124 = 264 bp ORF: 87 amino acids = 9.5 kDa |
| ORF: 6338 .. 6601 = 264 bp ORF: 87 amino acids = 8.9 kDa |
| ORF: 7190 .. 72 = 501 bp ORF: 166 amino acids = 17.5 kDa |
| ORF: 4855 .. 5079 = 225 bp ORF: 74 amino acids = 8.5 kDa |
| ORF: 5443 .. 5781 = 339 bp ORF: 112 amino acids = 12.6 kDa |
| ORF: 6358 .. 7317 = 960 bp ORF: 319 amino acids = 34.1 kDa |
| ORF: 69 .. 296 = 228 bp ORF: 75 amino acids = 7.9 kDa |
| ORF: 3270 .. 3536 = 267 bp ORF: 88 amino acids = 9.2 kDa |
| ORF: 5685 .. 6302 = 618 bp ORF: 205 amino acids = 21 |