Hello. We want to try TetR\CymR inducible system for our cells. I've read a lot about these systems but still unsure is the position of operator site in a repression configuration is important.
Usually, people put TetO\CuO site right downstream of promoter, but what if it will be positioned further downstream? Hypothetically polymerase should stuck at this site (if it's bound by CymR\TetR) and gene expression should be disrupted. But I did not find if anyone did like that.
Example:
Promotor | Gene1 | T2A | TetO\CuO | Gene2
Will Gene2 be transcribed only under induction?
Reference:
Article How to Choose the Right Inducible Gene Expression System for...
I am talking about repressor configuration - Fig 1A\2A
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