Yes, you can do this easily. Just install multisite and import on main site.
Process:
You have running mydomain.com. Now keep backup this website.
Now,
1. install website on multi-site
2. Import complete webiste on main domain mydomain.com
3. Set permalink as per your requirement on main site.
4. Add another site from network. Export theme setting and import setting on another site.
Best of Luck.
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