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 &# |