Gt(ROSA)26Sor^{tm1(CAG-Brainbow2.1)Cle}

A targeting vector containing (from 5' to 3') a strong CAGG promoter, a loxP site, a PGK-Neo-pA cassette (serving as a transcriptional roadblock), and the Brainbow 2.1 construct. This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. The Brainbow 2.1 construct was designed by Drs. Jeff Lichtman and Joshua Sanes (Harvard University) with four fluorescent protein sequences uniquely positioned in a tandem fashion and delimited by loxP sites in opposite orientation. Specifically, this Brainbow 2.1 coding region is composed of two adjacent floxed head-to-tail tandem dimers. The first head-to-tail dimer contains a loxP site and humanized Renilla GFP (hrGFPII; with nuclear localization signal plus polyA sequence) in forward orientation and a loxP site and monomeric EYFP (mYFPA206K plus polyA sequence) in reverse orientation. The second head-to-tail dimer contains a loxP site and tdimer2 RFP plus polyA sequence in forward orientation, and a loxP site and mCerulean CFP (with membrane tethering palmitoylation sequence plus polyA sequence) in reverse orientation. A single frt site is located at the 3' end of the Brainbow 2.1 construct. (J:125961, J:164644)