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. 2005 Dec 14;33(22):7039–7047. doi: 10.1093/nar/gki1009

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© The Author 2005. Published by Oxford University Press. All rights reserved

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General strategy to generate a programmed restriction enzyme. The single chain variant of PvuII is covalently linked via a (CH₂)₆- or (CH₂)₁₂ - linker to a TFO using GMBS, a bifunctional chemical cross-linker specific for amino and sulfhydryl groups. The final constructs, scPvuII-_C6-TFO and scPvuII-_C12-TFO, have a bipartite recognition site: the PvuII recognition site and the TFS.