PrecisionShuttle™ mammalian expression vector with TurboGFP and puromycin resistance markers, for fusing an ORF to C-terminal c-Myc and FLAG® tags.
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Sticky ends from different AlwNI sites may not be compatible.
Efficient cleavage requires at least two copies of the SacII recognition sequence.
Sticky ends from different BstEII sites may not be compatible.BstEII is typically used at 60°C, but is 50% active at 37°C.
The 1-base overhangs produced by PflFI may be hard to ligate.Sticky ends from different PflFI sites may not be compatible.
The 1-base overhangs produced by Tth111I may be hard to ligate.Sticky ends from different Tth111I sites may not be compatible.
BsrGI is typically used at 37°C, but is even more active at 60°C.
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
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.
PaeR7I does not recognize the sequence CTCTCGAG.
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.