| 31 May 25 |
r07        | Charlotte EaglesCharlotte Eag | 2 - 1 | Wake FCWake FC |
| 31 May 25 |
r07        | Reading UnitedReading Unite | 1 - 3 | Real Central New JerseyReal Central |
| 31 May 25 |
r07        | Ocean City Nor'eastersOcean City No | 2 - 0 | Delaware FCDelaware FC |
| 31 May 25 |
r07        | Lionsbridge FCLionsbridge F | 2 - 2 | Northern Virginia FCNorthern Virg |
| 31 May 25 |
r07        | Birmingham Legion 2Birmingham Le | 1 - 0 | Apotheos FCApotheos FC |
| 31 May 25 |
r07        | Fort Lauderdale UnitedFort Lauderda | 1 - 0 | St. Petersburg FCSt. Petersbur |
| 31 May 25 |
r07        | Athens UnitedAthens United | - | Columbus UnitedColumbus Unit |
| 31 May 25 |
r07        | SC United BantamsSC United Ban | 2 - 1 | Tobacco RoadTobacco Road |
| 31 May 25 |
r07        | Charlottesville BluesCharlottesvil | 0 - 1 | Virginia Beach UnitedVirginia Beac |
| 01 Jun 25 |
r07        | Westchester FlamesWestchester F | 1 - 1 | Ironbound SCIronbound SC |
| 01 Jun 25 |
r07        | Little Rock RangersLittle Rock R | 5 - 1 | Red RiverRed River |
| 01 Jun 25 |
r07        | Patuxent Football AthletiPatuxent Foot | 1 - 1 | Virginia MaraudersVirginia Mara |
| 01 Jun 25 |
r07        | Hill Country LobosHill Country | 3 - 5 | Lonestar SCLonestar SC |
| 01 Jun 25 |
r07        | Twin City ToucansTwin City Tou | 0 - 0 | AC Houston SurAC Houston Su |
| 01 Jun 25 |
r07        | Denton DiablosDenton Diablo | 1 - 0 | Mississippi BrillaMississippi B |
| 01 Jun 25 |
r07        | McKinney ChupacabrasMcKinney Chup | 3 - 5 | Hattiesburg FCHattiesburg F |
| 01 Jun 25 |
r07        | Dothan UnitedDothan United | 2 - 1 | Southern Soccer AcademySouthern Socc |
| 01 Jun 25 |
r07        | Tacoma StarsTacoma Stars | 0 - 0 | Portland BangersPortland Bang |
| 01 Jun 25 |
r07        | Snohomish UnitedSnohomish Uni | 1 - 3 | Midlakes UnitedMidlakes Unit |
| 01 Jun 25 |
r07        | West Seattle JunctionWest Seattle | 4 - 5 | FC OlympiaFC Olympia |
| 01 Jun 25 |
r07        | Long Island Rough RidersLong Island R | 2 - 0 | Morris EliteMorris Elite |
| 03 Jun 25 |
r07        | Southern California EagleSouthern Cali | 3 - 1 | Stars FCStars FC |
| 03 Jun 25 |
r07        | CISACISA | 0 - 1 | Flatirons RushFlatirons Rus |
| 03 Jun 25 |
r07        | Lansing City FootballLansing City | 1 - 2 | Union FC MacombUnion FC Maco |
| 04 Jun 25 |
r07        | Peoria CityPeoria City | 6 - 0 | Springfield FCSpringfield F |
| 04 Jun 25 |
r07        | Colorado StormColorado Stor | 1 - 0 | Albion SC ColoradoAlbion SC Col |
| 04 Jun 25 |
r07        | West Virginia UnitedWest Virginia | 0 - 3 | Fort Wayne FCFort Wayne FC |
| 04 Jun 25 |
r07        | Kings Hammer FCKings Hammer | 0 - 3 | Lexington SC IILexington SC |
| 04 Jun 25 |
r07        | Rochester FCRochester FC | 2 - 1 | RKC Third CoastRKC Third Coa |
| 04 Jun 25 |
r07        | Midwest UnitedMidwest Unite | 2 - 0 | Flint City BucksFlint City Bu |
| 05 Jun 25 |
r07        | Santafé WanderersSantafé Wande | 3 - 4 | Des Moines MenaceDes Moines Me |
| 05 Jun 25 |
r07        | Marin FC LegendsMarin FC Lege | 0 - 3 | San Francisco City FCSan Francisco |
| 05 Jun 25 |
r07        | Monterey Bay FC 2Monterey Bay | 0 - 1 | Academica SCAcademica SC |
| 05 Jun 25 |
r07        | Steel CitySteel City | 2 - 0 | Cleveland ForceCleveland For |
| 06 Jun 25 |
r07        | Northern IndianaNorthern Indi | 4 - 1 | Kalamazoo FCKalamazoo FC |