A targeting vector was designed to insert a FLAG/bio epitope tag and a frt-flanked neomycin resistance (neo) cassette at the endogenous Srf stop codon and upstream of the 3' UTR.

The construct was electroporated into 129S4/SvJae-derived J1 embryonic stem (ES) cells and correctly targeted ES cells were injected into blastocysts. The resulting chimeric mice were bred to outbred Swiss Webster mice. Offspring were bred with Tg(ACTFLPe)9205Dym transgenic mice to delete the neo cassette. The resulting Srf-flbio mice were maintained on a Swiss Webster background.

The Gt(ROSA)26Sor^{tm1.1(birA)Mejr} allele was created using an HA (hemagglutinin) tagged bacterial birA, bifunctional protein (biotin ligase), gene inserted into exon 3 of the rabbit beta-globin gene. The entire birA ORF, with the rabbit beta globin splice acceptor and transcription termination sequences were excised to generate a targeting vector that also contained a floxed PGK- puromycin selection cassette. This targeting vector was inserted into the first intron of the Gt(ROSA)26Sor gene. The construct was electroporated into 129P2/OlaHsd derived IB10 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to FVB/N mice. The mice were intercrossed for 3 generations before being crossed to a Cre recombinase expressing strain (on an unknown genetic background) to remove the loxP-flanked PGK-puromycin selection cassette. A single loxP site remains. The Cre transgene was removed by breeding.

Mice homozygous for both alleles mice were maintained by the Cardiovascular Development Consortium (CvDC), an NIH-funded consortium within the Bench-to-Bassinett (B2B) program.

Upon arrival, mice were bred to FVB/NJ inbred mice for at least one generation to establish the colony.

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