Just wondering how you obtained you 1.5 kb orf, cut from another vector or from PCR and how you prepared your vector.
We cut our ORF from another vector. we both cut our insert and vector with SacI and SalI, we did the restriction seperately. First, we cut with SacI and run on LMP agarose and isolate samples and later we cut with SalI and run on gel again and isolate it from the gel. Then set up several ligation as I have told.