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. Author manuscript; available in PMC: 2012 Jul 15.
Published in final edited form as: J Neurosci Methods. 2011 Apr 8;199(1):1–9. doi: 10.1016/j.jneumeth.2011.03.028

Figure 2. Preparation of the DNA constructs.

Figure 2

A) Structure of the Tn5 transposon: The black segments: 19-bp mosaic end (ME) sequences required for transposition. The orange segment on left: a full length sequence for venus. The pink segment: the sequence for Kanamycin resistance. The orange segment on the right: C-terminal sequence of venus. B) Transposon reaction: The transposase inserts the transposon into the targeted vector at various locations. C) The products of the transposon reaction are tested by looking for fluorescence in transfected HEK293 cells. The ones with wrong orientation, out-of-frame or out-of-coding-region do not produce venus. D) The constructs with correct insertions were cut by either AsiS1 or AsC1 and then ligated. The deletions generated constructs containing either the N-terminal segment or the C-terminal segment of the venus.