making reading frame - (Apr/07/2006 )

Hi i want to clone a gene for expression. I have a gene sequence and I want to know that to make a reading frame I have to make it in 5' to 3' sequecne and make its reading frame in some tools. If yes ?????

I have my gene sequence from 5' to 3' direcation and I have make its reading frame its reading frame was 3
but when i remove some sequece from 5' direction now its completely in readinf frame 1
and i have attached enzyme site at its 5' point and make its reading frame its again in reading frame and it was 1 reading frame and i used
9 vector sequece or vector sequence multiple of 3 then its reading frame is ok if i donot use vector sequecne multiple of 3 then its not in the reading frame. I have to clone my gene into pET32 a +
how much vector sequece i have to take ever multiple of 3 or u can take any no of vector sequece and it shold ever be multiple of 3 because if i take multple of 3 then my frame is ok in pET32 a+ if not multiple of 3 then its not in reading frame.
please correct me
best regards

Your start codon defines your coding sequence. You can add what ever you want to the 5' end of the sequence between the promoter and start seqence as long as there are no other start codons in that sequence. It's bad technique to include a lot of sequence between the promoter and start, but sometimes it can't be avoided.
Good Luck