The article describes a comprehensive atlas of the mid-gestational human brain, including *de novo* reference atlases, *in situ* hybridization, ultra-high resolution magnetic resonance imaging (MRI), and microarray analysis of highly discrete laser microdissected brain regions. The study reveals transcriptional differences between different proliferative and postmitotic layers of the developing cerebral cortex, with laminar signatures reflecting cellular composition and developmental processes. Species differences in gene expression are observed, particularly in the subplate, and minimal differences are found between the inner and outer subventricular zones. Fronto-temporal gradients are observed in both germinal and postmitotic cortical layers, with enrichment in the frontal lobe. Many neurodevelopmental disorder and human evolution-related genes show patterned expression, potentially underlying unique features of human cortical formation. These data provide a rich resource for understanding human brain development.The article describes a comprehensive atlas of the mid-gestational human brain, including *de novo* reference atlases, *in situ* hybridization, ultra-high resolution magnetic resonance imaging (MRI), and microarray analysis of highly discrete laser microdissected brain regions. The study reveals transcriptional differences between different proliferative and postmitotic layers of the developing cerebral cortex, with laminar signatures reflecting cellular composition and developmental processes. Species differences in gene expression are observed, particularly in the subplate, and minimal differences are found between the inner and outer subventricular zones. Fronto-temporal gradients are observed in both germinal and postmitotic cortical layers, with enrichment in the frontal lobe. Many neurodevelopmental disorder and human evolution-related genes show patterned expression, potentially underlying unique features of human cortical formation. These data provide a rich resource for understanding human brain development.