The 2.5 kb SalI/BglII fragment contains internal sites for AvaI, PvuI, EcoRI, XhoI, BstEII (one site each) and PstI, SacI (2 sites each). Restriction digests of the clone give the following sizes (kb): EcoRI--23, 16; BglII--17, 12, 6.0, 4.7, 3.9; SalI--18, 15, 11. This pattern differs from the information supplied by the depositor. The hygromycin B and G418 resistance structural genes and control elements can be isolated on the 7.5 kb BglII fragment or the 2.5 kb SalI/BglII fragment. The 1.5 kb BglII/SacI fragment contains hygR. The 1.65 EcoRI/SalI fragment contains G418R. The 7.5 kb BglII fragment contains a prokaryotic replicon and can be self-ligated to form plasmid pSC701. This is a vector with easily isolated hygromycin B and G418 resistance structural genes and control elements for use in constructing other plasmids. |
