pETDuet-1
Bacterial vector for the co-expression of two genes.
Sequence Author: MilliporeSigma (Novagen)
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After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility. |
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| * Blocked by Dam methylation. |
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| * Blocked by Dam methylation. |
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Efficient cleavage requires at least two copies of the SgrAI recognition sequence. |
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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 BstAPI 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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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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Sticky ends from different PpuMI sites may not be compatible. |
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Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present. This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.Sticky ends from different Bpu10I sites may not be compatible. |
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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 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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BsrGI is typically used at 37°C, but is even more active at 60°C. |
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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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FseI gradually loses activity when stored at -20°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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PaeR7I does not recognize the sequence CTCTCGAG. |
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Sticky ends from different BlpI sites may not be compatible. |
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Sticky ends from different DraIII 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. |
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Sticky ends from different BsaI sites may not be compatible.BsaI can be used between 37°C and 50°C. |
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Sticky ends from different BglI sites may not be compatible. |
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Sticky ends from different AlwNI sites may not be compatible. |
| pET Upstream Primer 16-mer / 63% GC 1 binding site 5358 .. 5373 = 16 annealed bases Tm = 57°C |
| DuetUP2 Primer 20-mer / 50% GC 1 binding site 189 .. 208 = 20 annealed bases Tm = 57°C |
| DuetDOWN1 Primer 20-mer / 50% GC 1 binding site 189 .. 208 = 20 annealed bases Tm = 57°C |
| T7 Terminator Primer 19-mer / 53% GC 1 binding site 448 .. 466 = 19 annealed bases Tm = 57°C |
| lacI 3931 .. 5013 = 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 3931 .. 5013 = 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 |