Yes to what Dan said above^^
With a Layout, you can add a Map Frame. This looks to one of your maps from the project and all the layers within that map. So yes, you will need to first configure your map with all its necessary layers, then add the map into your Layout using Map Frame (Insert tab).
You can add layers into your map by using the Add Data button on the ribbon, or clicking and dragging from the Catalog.
Thanks,
Gillian