Hello Benjamin.

That should be found only by experimental evidences, but not by general knowledge of molecular biology. However, I guess that a local conformation and concentrations of binding components in the poly(A) complex would be limiting factors for transcription step.

Regards.

Why do you have 2 polyA sequences? As long as the 3' polyA sequence is not affecting the 5 ' polyA sequence then you should have no problems. The stability of the message should also be ok.

Could you remove one of the polyA sequences ?

Thanks,

Angel

Thanks Sang Ho and Angel. I did end up removing the second polyA sequence, although it cost me an extra few days :). More than anything, I was just wondering if anyone had looked at expression in this setting off of a viral vector (which must package the genome and therefore repeating polyA sequences could be disruptive.