I am using pcDNA myc his (-) 3.1 vector for cloning. in the original vector, it has around 50bp spacer between the stop codon and BGH terminator. But i want to insert a stop codon before the original stop codon of the vector. then the space between stop codon and the terminator will be around 150 bp. Would it affect to the downstream expression efficiency?. I hope some suggestions.
Hi Nipuna,
I hope there will be no issue due to the distance between the stop codon and the terminator sequence being around 150 bp. In bacterial genomes, the distance between the stop codon and the terminator sequence can be zero to a few hundred bp long. More often it is less than 50 bp. Termination within 100 bp often works efficiently. It can be verified by just conducting the expression efficacy after cloning. I expect there will be no significant issue.
Thank you,
Amal.
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