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. 2019 Sep 16;9:13330. doi: 10.1038/s41598-019-49783-w

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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ZsGreen transgene. (A) The transgene construct contains a chicken β-actin promoter with a CMV enhancer element followed by a chimeric intron. The floxed-STOP region contains loxP sites flanking 3 tandem SV40 polyadenylation signal sequences. Downstream of the floxed-STOP cassette is the ZsGreen gene sequence (originally derived from a Zoanthus sp. reef coral) followed by a bovine growth hormone polyadenylation signal. Upon Cre recombinase-mediated recombination, the loxP sites are recombined resulting in the excision of the 3 tandem SV40 polyadenylation signals. This event allows expression of the ZsGreen gene. (B) PCR amplification to demonstrate removal of the floxed STOP region from the ZsGreen-containing transgene. Lane 1: ZsGreen transgenic animal; Lane 2: NCre transgenic animal; Lane 3: double transgenic animal. Upper panel: results of amplification with primers flanking the floxed STOP region. Lower panel: results of amplification with primers that detect the Cre recombinase gene. A molecular size standard is shown to the left of the image. For reference, the uncropped image is included as Fig. S1.