Thanks for the sequence
-- The plasmid has Lac O and it is functional.
The reason that you don't need to induce it is that E. coli typically only makes a few copies of the lac repressor per cell. With a high-copy plasmid, there are too many operator sites and most of them are unoccupied.
I put this plasmid into XL1-blue, which has the constitutive repressor (LacI^q) on an F'. The GFP is repressed in that strain and should be inducible with IPTG.
I'm planning to test that this week.
I had one of my students do the transformation and we didn't have any amp/tet plates for it (The F' is maintained by Tet^r). Out of a few hundred not-so-green colonies, there were 3 bright green ones that had lost the F' and therefore the constitutive repressor.
We're planning a series of labs looking at gene regulation and having the students "discover" the rare bright green colonies in the absence of inducer, then plan experience to figure out why. Let me know if anybody wants the procedure once we test it.
-tony
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