Hi guys,
This might be a dumb question, but I can't locate the terminator sequences for the lacZ gene in the pUC19 plasmid. Apparently none of the maps in the great and mighty Internet (aka Google) for this plasmid have the terminator sequences annotated. Even when I BLAST the sequences from the T1 and T2 terminators from E. coli to the plasmid sequence, I get zero results.
Is there a way to locate them in the plasmid sequence? I tried to use NUPACK to predict the formation of hairpins in certain regions of the plasmid, but I can't get the coordinates for the hairpins from this software. Also, does anybody have any idea why those sequences are not annotated in the plasmid? pUC19 is a vastly used plasmid, and I can't think of any reason why all the maps I've found does not have the terminators.
There is still a possibility that it does not contain terminator after lacZ'.
Hi guys, thanks for the help!
Priyanka, I followed your suggestion and used the RibEx tool to locate the terminator sequences. Turns out that Danila was right: there is a terminator after the bla gene in the pUC19 plasmid, but apparently no terminator after the lacZ gene. That makes sense as the two genes are located in the same strand and I believe the absence of a terminator after the lacZ gene would increase the probability of the two genes to be expressed.
WebDSV, an online molecular cloning software, can auto-detect miscellanous DNA sequence elements.
http://www.molbiotools.com/WebDSV/
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