Igs7^{tm218(tetO-rho*/mNeonGreen,CAG-mRuby3,-tTA2)Tasic}

The targeting vector has a Tet response element (tetO), loxP-STOP-loxP cassette and a Ace2N-4AA-mNeonGreen-WPRE sequence that is followed by a CAG promoter, Lox2272-STOP-Lox2272 cassette, mRuby3, and IRES-tTA2-WPRE sequence - all inserted into the T1 TIGRE locus (an intergenic region on chromosome 9 [between AB124611 and Carm1] determined to have low basal transcriptional activity and allow high cre and/or Tet inducibility). Ace2N (rho*)-4AA-mNeonGreen is an indicator variant of Acetabularia acetabulum-derived rhodopsin (Ace) with an inactivated proton pump (Ace2N), with four extra amino acids in a linker domain that fuses to mNeonGreen fluorescent protein to enable voltage-sensitive fluorescence resonance energy transfer (FRET). Of note, the Ai218 allele was designed with an IRES upstream of the tTA2 to reduce/eliminate leakiness in the absence of cre recombinase. (J:101977)