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. 2017 Feb 16;45(10):e89. doi: 10.1093/nar/gkx113

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© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — In vitro transposition assay (in vitro hop assay). A donor plasmid contains a selection cassette (kanamycin resistance, Kan^R) flanked with the inverted repeats (IR), which are recognized by transposase. Upon addition of divalent ions Mg²⁺ or Mn²⁺in vitro, purified transposase excises the selection cassette and integrates it into the target plasmid. Purified products of in vitro transposition can be analyzed by transfection into Escherichia coli DH10B cells and selection in the presence of kanamycin. The donor plasmid contains a conditional origin of replication oriR6K and will not propagate in the destination strain; only the kanamycin cassette containing target plasmids are maintained.