Hi all,
I am integrating a two gene pathway into the genome of E.coli. I have a good locus in which to integrate this and I have selected a good promoter. However, I am wondering do I need to incorporate a terminator after the gene also? This may be a silly question but I am new to this technique and prefer to ask stupid questions rather than do stupid mistakes.
Best
John
Yes, I believe you should include a transcription terminator into your construct. If you are relying on a native E. coli promoter, that is well studied, then it may already have a native transcription terminator that you can rely on. However, if not or there no characterized terminator for your locus, then I recommend engineering a terminator in your construct. I recommend terminators tR' or tR2 (which is smaller than tR' if you are restricted on size) for your construct.
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