质粒测序两个基因均正确，双酶切，只有一个可以切开，求帮助！！...

两个基因大小分别230kb和450kb: are they in differerent vectors? What kinds of vectoris it if they do in the vector? Are you sure that the two genes are in several hundred kb, not bp? Do you have any gel picture?

用的一样的载体，都是pGEM，是230bp和450bp，没有拍照，下次拍了上传一下。