Previously, we proposed the Antigen-mediated genetically modified cell amplification (AMEGA) system, which can selectively amplify gene-transduced cells without antibiotic selection. While the original AMEGA system employed two vectors encoding a pair of antibody/receptor chimeras and a gene of interest, the use of two virions resulted in low co-transduction efficiency and time-consuming selection. Here we show an improved AMEGA system, where the three genes are linked in tandem with internal ribosomal entry sites in a single vector. We used gp130 chimeras whose extracellular domain was replaced with either V-H or V-L region of anti-hen egg lysozyme (HEL) antibody HyHEL-10 and D2 domain of erythropoietin receptor. The constructed vector was retrovirally transduced to IL-3-dependent Ba/F3 cells followed by HEL selection in the absence of IL-3. The resultant transduction efficiency increased by similar to100-fold compared to the previous two-vector system, which results in a much shorter selection period.