Hi everybody,
I have two plasmids:
- One expressing the CRY2-CIB1 optogenetic system fused to the TetA transcription factor (TetA is reconstituted upon stimulation with blue light). This one has a constitutive promoter.
- The other one expresses a reporter fluorophore under the Tet promoter.
Does someone have any advice on how to join both in a single plasmid? Is it possible to have a plasmid with two different promoters?
I am trying to avoid co-transfection of the two, because the efficiency is obviously lower than by having a single construct.
Thanks to anybody that can help!
Nearly all plasmids have multiple promoters (for E. coli) so this should not be a problem.
Thanks for your answer! These are for mammalian expression.
I suppose there exists a proper design principle (e.g. 1st promoter - 1st coding sequence - stop codon - 2nd promoter - 2nd coding sequence) that ensures correct processing, but I haven't found a clear answer to this.
There does not need to be any order since each promoter generates its own transcript and are therefore transcriptionally independent. They don't even need to be oriented the same way.
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