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. 2019 Jan 11;9(3):737–748. doi: 10.1534/g3.118.200975

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Copyright © 2019 Williams et al.

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

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Strategies for generating conditional tagged variants of a Drosophila Rab3 synaptic vesicle marker. A) top: genomic exon structure of Drosophila Rab3. middle: strategy 1 involves incorporating a B2 recombinase target (B2RT)-flanked transcription STOP cassette upstream of exon 2 and either a 2XHA epitope tag, or a GFP or mCherry fluorescent protein tag, at the amino-terminus of Rab3. Prior to excision of the transcription STOP cassette, the tagged variants of Rab3 will not be expressed. bottom: after B2 recombinase expression in neurons of interest, the tagged variants of Rab3 will be expressed in neurons in which the STOP cassette has been excised. B) top: strategy 2 involves paired FRT and F3 FLP recombinase target sites flanking an inverted exon 2 containing 2XHA or 3XFLAG amino-terminal epitope tags. bottom: after FLP recombinase expression in neurons of interest and an odd number of inversions followed by an excision, the tagged variants of Rab3 will be stably expressed. Note that FLP recombinase is functional with FRT pairs and F3 pairs but is not functional in an FRT-F3 combination.