Well, The most easily available vector is of course the kits. But if you are
that adverse to using them, then utilize the cutting ability of Xcm I, which
recognizes CCANNNNNNNNNTGG, and leaves a single 3' base overhang.
An appropriately designed oligonucleotide with 2 XcmI sites put in vectorofchoice will generate
a non self ligating linear band with the 3' T overhang.
This is how we did it before the kits came out.
Dept of Microbiology
University of Reading