Primary Identifier | MGI:4947243 | Allele Type | Targeted |
Attribute String | Reporter | Gene | Gt(ROSA)26Sor |
Transmission | Germline | Strain of Origin | (129S6/SvEvTac x C57BL/6NCrl)F1 |
Is Recombinase | false | Is Wild Type | false |
molecularNote | The Rosa-CAG-LSL-Synaptophysin-tdTomato-WPRE targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), an FRT site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), a Synaptophysin-tdTomato fusion protein sequence, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a bovine growth hormone polyA signal, and an attB/attP-flanked PGK-FRT-Neo-polyA cassette. To create the Synaptophysin-tdTomato fusion protein sequence, a mouse synaptophysin cDNA sequence (encoding a 308 amino acid transcript with all but the first six amino acids of the full-length protein) was fused in-frame via a six nucleotide linker to the amino terminus of the tdTomato sequence (a non-oligomerizing DsRed fluorescent protein variant with a 12 residue linker fusing two copies of the protein (tandem dimer)). PhiC31-mediated recombination removed the neo cassette. |