Hi All,

I am designing a vector carrying a bicistronic transgene with two promoters that will be integrated into the host's genome. I am thinking of adding three insulators (the same type): two flanking the whole transgene and one separating the two promoters, for their barrier and enhancer blocker activity, respectively.

Do insulator sequences require a separation of some nucleotides from promoters and polyA tails, or can I place them straight before or after them?