I am trying to express m6A protein and a fluorescence one RFP in the piggyback vector.

I was thinking first of using the P2A to separate between the m6A and the RFP protein but then I was afraid of the residual P2A in the m6A if it would affect the localisation or the interaction at the C terminus, then the second choice would be IRIS, although the expression casste in the Piggybac itself has another IRIS followed by the selection marker

so the cassette would be as follows

m6A IRIS RFP IRIS puromycin

Is that ok or I would have a problem with the expression of the proteins

If there is please can you suggest other thoughts?

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