pTandem-1
Mammalian vector for the co-expression of two genes from a bicistronic mRNA using the CMV promoter and an IRES.
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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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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Efficient cleavage requires at least two copies of the NmeAIII recognition sequence. Sticky ends from different NmeAIII sites may not be compatible.For full activity, add fresh S-adenosylmethionine (SAM). |
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Efficient cleavage requires at least two copies of the BpmI recognition sequence. Sticky ends from different BpmI sites may not be compatible.After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.BpmI quickly loses activity at 37°C. |
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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, BseYI can remain bound to DNA and alter its electrophoretic mobility. |
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Sticky ends from different DrdI 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. |
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| * Blocked by Dam methylation. |
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Sticky ends from different BtgZI sites may not be compatible.After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.BtgZI is typically used at 60°C, but is 75% active at 37°C. |
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Efficient cleavage requires at least two copies of the BsgI recognition sequence. Sticky ends from different BsgI sites may not be compatible.For full activity, add fresh S-adenosylmethionine (SAM). |
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Efficient cleavage requires at least two copies of the SacII recognition sequence. |
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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 XcmI may be hard to ligate.Sticky ends from different XcmI 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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PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations. |
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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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BssHII is typically used at 50°C, but is 75% active at 37°C. |
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