Good morning.
I am trying to obtain information about the sequence or at least a restriction map of Carolina's pGREEN plasmid. I am supposed to cut it with BbsI, but when I try to make a virtual digest using pGREEN sequences from NCBI I don't get any site or only one.
We have been told to use specifically that plasmid and that restriction enzyme, so I would be very grateful if anyone has information about the plasmid.
Thanks.
http://www.snapgene.com/resources/plasmid_files/plant_vectors/pGreen/
I don't see BbsI as a cutter in the plasmid on the above map. I also found a sequence of pGreen, and although it had some ambiguous nucleotides, I ran it through NEB cutter and BbsI was listed as a non-cutter.
With that said, if your protocol tells you to digest with BbsI, it's possible that they engineered a BbsI restriction site into pGreen which is why I can't find it listed as a cutter.
It's worth ~1.5hrs of your time to run a gel with undigested plasmid and BbsI digested plasmid to see if you actually get single cut plasmid or supercoiled.
Hope this helps.
Carolina's pGREEN isn't the snapgene plant vector pGreen. https://groups.google.com/forum/#!topic/diybio/8zru-2gdk9w
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