pHTN HaloTag CMV-neo
Mammalian expression vector with a traditional MCS and ampicillin and G418 resistance markers, encoding a cleavable N-terminal HaloTag®.
Sequence Author: Promega
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PciI is inhibited by nonionic detergents. |
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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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After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility. |
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Sticky ends from different BstXI 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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Efficient cleavage requires at least two copies of the NaeI recognition sequence. |
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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 PluTI recognition sequence. |
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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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Sticky ends from different PflMI sites may not be compatible. |
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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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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 PasI 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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PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations. |
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Efficient cleavage requires at least two copies of the SgrAI recognition sequence. |
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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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Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.Sticky ends from different AccI sites may not be compatible. |
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PaeR7I does not recognize the sequence CTCTCGAG. |
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Efficient cleavage requires at least two copies of the SacII recognition sequence. |
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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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ApaI can be used between 25°C and 37°C. |
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Sticky ends from different BlpI sites may not be compatible. |
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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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Efficient cleavage requires at least two copies of the SfiI recognition sequence. Sticky ends from different SfiI sites may not be compatible. |
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Efficient cleavage requires at least two copies of the NarI recognition sequence. |
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| HaloTag® 1067 .. 1957 = 891 bp 297 amino acids = |