pcDNA3.2 V5-DEST
Gateway® destination vector for high-level expression of C-terminally V5-tagged proteins in mammalian cells.
Sequence Author: Thermo Fisher (Invitrogen)
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PciI is inhibited by nonionic detergents. |
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Sticky ends from different PfoI sites may not be compatible. |
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Efficient cleavage requires at least two copies of the RsrII recognition sequence. Sticky ends from different RsrII sites may not be compatible.For full activity, add fresh DTT. |
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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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Efficient cleavage requires at least two copies of the PluTI recognition sequence. |
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Efficient cleavage requires at least two copies of the NarI recognition sequence. |
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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 CsiI sites may not be compatible. |
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| * Blocked by Dcm methylation. Sticky ends from different SexAI sites may not be compatible. |
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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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Sticky ends from different BstXI sites may not be compatible. |
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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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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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ApaI can be used between 25°C and 37°C. |
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Efficient cleavage requires at least two copies of the SacII recognition sequence. |
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Sticky ends from different DraIII sites may not be compatible. |
| AmpR 6715 .. 7575 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 6715 .. 7506 = 792 bp 263 amino acids = |