Schematics of fluorescent mGAT1 construct designs and primary sequence alignments of the regions that are fused with XFP. In each schematic, the mGAT1 peptide is illustrated as a black bar, and TMs are marked as superimposed numbered boxes. TM12 is highlighted in magenta. Linkers deriving from cloning vector are shown in red, and the XFP moiety is shown as a cyan and yellow box. Additional hGAT1 sequence is shown as a dark blue box. #, a variable number of residues depending on the construct design. All schematics and their internal features are scaled according to sequence length. The three classes of florescent mGAT1 fusions, C-terminal additions, C-terminal fusions, and N-terminal additions are displayed with their names to the left. For the fusions of XFP to the mGAT1 C terminus, only the last 14 residues of the fluorophore and any appended hGAT1 or other sequence are displayed. For the mGAT1 C-terminal XFP insertions, the regions immediately adjacent to the insertion site and the first four and last four residues of the XFP are displayed. Displayed GAT1 C-terminal sequences are numbered with the terminal residue being P(0); upstream sequence positions are negative. Other important highlighted regions include: fluorophore sequence (GFP in green; XFP in light blue), the terminal hydrophobic residues of mGAT1₀GFP and mGAT1XFP* (purple), the endogenous GAT1 class II PDZ–interacting motif -AYI (dark green), mouse or hGAT1 TM12 sequence (magenta), nonclassical endocytic motifs homologous to FREKLAYAIT in mDAT (conserved residues underlined) (Holton et al., 2005; Boudanova et al., 2008), RL or RI ER export motif (bold) (Farhan et al., 2007), and VMI or VMV (bold) ER–Golgi intermediate compartment export motif (Farhan et al., 2008). XFP sequence position in XFP15mGAT1 constructs is displayed below the sequence in blue, and mGAT1 numbering is in black. Linker sequence between XFP and mGAT1 is shown in red.