Your plasmid is small and does not seem to have a lot of dead space. I would suggest you either put it downstream of the CYC1 term or upstream of the TEF promoter. Downstream of CYC1 term is preferable since it eliminates the possibility of interfering with the expression from the TEF promoter.

Thanks Hanna!

Do you think adding a few nucleotide at both sides of the ARS sequences will be helpful to create more space and further minimize the possibility of interfering with other events?

It one of those where you have to try and see, I cannot tell you for sure that adding a few bases will change anything, especially since it might not even impact what you are looking for. I suggested downstream of the CYC1 term since it the least likely position to affect either your gene/selection marker expression since it is distant to your gene and the 3’UTR of the selection marker should theoretically have been formed at that point.

Thank you for the explanation, this is really helpful!

Good luck to me and I wish you success too :)