Hello,
We have this older plasmid YEpTOP2GAL1 for expression in S. cerevisiae. We don't have the plasmid sequence which would be necessary to design primers or new cloning constructs. Does any one have the sequence for YEpTOP2GAL1? I can't find it in any references or sites like Addgene.
Thanks!
References - PMID: 24124932, PMID: 23129636
Hello,
Getting an exact sequence for this plasmid would be a difficult since at least two parts of it where constructed by BAL digestion and the authors of the original paper that constructed the plasmid did not describe in details. However they indicate that the majority of the backbone comes from YEP24 (not the gal1 promoter) and a schematic diagram of YEpTOP2GAL1 can be found here:
Article DNA supercoiling in vivo
I suggest you compare the schematic with the YEP24 map from this link:
https://www.snapgene.com/resources/plasmid-files/?set=yeast_plasmids&plasmid=YEp24
take the common sites and sequence from there (for example the URA3 marker is closest to the TOP2 insert and hence you can design accordingly). A couple of good sequencing reaction will help you in inferring the entire sequencing of the plasmid.
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