To functionally validate our αβTCR cloning vector backbone we chose the well-characterized CD8+ T-cell clone 14/35 directed against mutant CDK4 (R24C) that has originally been established via an autologous mixed lymphocyte-tumor culture (MLTC) using the CDK4R24C-positive melanoma-cell line SK29-MEL.1 as stimulator of blood-derived T cells [12]. Here, CD8A is linked to melanoma.