I need the DNA sequence pCP13 cosmid vector. I know that pCP13 was derived from pLAFR1 and have the DNA sequence for pLAFR1 but I don't know the sequence of the kanamycin resistance gene (or where it came from) and I need this information. I would appreciate any insight about pCP13. Thank you!
that is a bit of a tricky one because the paper in which this plasmid was used indicates that they obtained it personally without the details of the construction being published. However, these plasmids all seems to be broad host range and I checked the restriction map of pCP13 in this ref:
Article Cloning of genes controlling alginate biosynthesis from a mu...
it indicated the presence of an xhoI and hindIII site within the kanR gene which is not very common in the common kanR marker in standard ecoli plasmids considering that hindIII is a standard site in most multiple cloning sites, therefore, you kanR marker must be a different one and is more likely present in broad host vectors. I find a plasmid that contains similar sites within the kanR gene which may likely to be the sequence you are looking for (I suggest you design primers and sequence the locus to confirm). The sequence is uploaded here I hope it helps.
Thank you for this information. This is an 'old' cosmid but I like to use it because it is reliable and stable. It was constructed in a time when the DNA sequence was not so readily available. I need the exact DNA sequence so I have made primers based on pLAFR1 (parent cosmid) and am sequencing the kanamycin resistance gene region and entering it into the NCBI database so everyone will have it.
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