diff --git a/extmod/modos.c b/extmod/modos.c
index 7f1e31fba..e7f7fc818 100644
--- a/extmod/modos.c
+++ b/extmod/modos.c
@@ -153,9 +153,6 @@ static const mp_rom_map_elem_t os_module_globals_table[] = {
     { MP_ROM_QSTR(MP_QSTR_putenv), MP_ROM_PTR(&mp_os_putenv_obj) },
     { MP_ROM_QSTR(MP_QSTR_unsetenv), MP_ROM_PTR(&mp_os_unsetenv_obj) },
     #endif
-    #if MICROPY_PY_OS_SEP
-    { MP_ROM_QSTR(MP_QSTR_sep), MP_ROM_QSTR(MP_QSTR__slash_) },
-    #endif
     #if MICROPY_PY_OS_SYNC
     { MP_ROM_QSTR(MP_QSTR_sync), MP_ROM_PTR(&mp_os_sync_obj) },
     #endif
@@ -170,6 +167,7 @@ static const mp_rom_map_elem_t os_module_globals_table[] = {
     #endif
 
     #if MICROPY_VFS
+    { MP_ROM_QSTR(MP_QSTR_sep), MP_ROM_QSTR(MP_QSTR__slash_) },
     { MP_ROM_QSTR(MP_QSTR_chdir), MP_ROM_PTR(&mp_vfs_chdir_obj) },
     { MP_ROM_QSTR(MP_QSTR_getcwd), MP_ROM_PTR(&mp_vfs_getcwd_obj) },
     { MP_ROM_QSTR(MP_QSTR_listdir), MP_ROM_PTR(&mp_vfs_listdir_obj) },
diff --git a/ports/nrf/mpconfigport.h b/ports/nrf/mpconfigport.h
index 0b9b667ee..48b58143a 100644
--- a/ports/nrf/mpconfigport.h
+++ b/ports/nrf/mpconfigport.h
@@ -151,7 +151,6 @@
 #define MICROPY_PY_OS_INCLUDEFILE   "ports/nrf/modules/os/modos.c"
 #define MICROPY_PY_OS_DUPTERM       (1)
 #define MICROPY_PY_OS_DUPTERM_STREAM_DETACHED_ATTACHED (1)
-#define MICROPY_PY_OS_SEP           (1)
 #define MICROPY_PY_OS_SYNC          (MICROPY_VFS)
 #define MICROPY_PY_OS_UNAME         (1)
 #define MICROPY_PY_OS_URANDOM       (MICROPY_HW_ENABLE_RNG)
diff --git a/ports/renesas-ra/mpconfigport.h b/ports/renesas-ra/mpconfigport.h
index 52effd64f..a23a2c106 100644
--- a/ports/renesas-ra/mpconfigport.h
+++ b/ports/renesas-ra/mpconfigport.h
@@ -120,7 +120,6 @@
 #define MICROPY_PY_OS_DUPTERM       (3)
 #define MICROPY_PY_OS_DUPTERM_BUILTIN_STREAM (1)
 #define MICROPY_PY_OS_DUPTERM_STREAM_DETACHED_ATTACHED (1)
-#define MICROPY_PY_OS_SEP           (1)
 #define MICROPY_PY_OS_SYNC          (1)
 #define MICROPY_PY_OS_UNAME         (1)
 #define MICROPY_PY_OS_URANDOM       (MICROPY_HW_ENABLE_RNG)
diff --git a/ports/stm32/mpconfigport.h b/ports/stm32/mpconfigport.h
index bd1ab671e..25fc9e11f 100644
--- a/ports/stm32/mpconfigport.h
+++ b/ports/stm32/mpconfigport.h
@@ -99,7 +99,6 @@
 #define MICROPY_PY_OS_DUPTERM       (3)
 #define MICROPY_PY_OS_DUPTERM_BUILTIN_STREAM (1)
 #define MICROPY_PY_OS_DUPTERM_STREAM_DETACHED_ATTACHED (1)
-#define MICROPY_PY_OS_SEP           (1)
 #define MICROPY_PY_OS_SYNC          (1)
 #define MICROPY_PY_OS_UNAME         (1)
 #define MICROPY_PY_OS_URANDOM       (MICROPY_HW_ENABLE_RNG)
diff --git a/ports/unix/variants/mpconfigvariant_common.h b/ports/unix/variants/mpconfigvariant_common.h
index 2e34055bf..cea039741 100644
--- a/ports/unix/variants/mpconfigvariant_common.h
+++ b/ports/unix/variants/mpconfigvariant_common.h
@@ -95,7 +95,6 @@
 #define MICROPY_PY_OS_INCLUDEFILE      "ports/unix/modos.c"
 #define MICROPY_PY_OS_ERRNO            (1)
 #define MICROPY_PY_OS_GETENV_PUTENV_UNSETENV (1)
-#define MICROPY_PY_OS_SEP              (1)
 #define MICROPY_PY_OS_SYSTEM           (1)
 #define MICROPY_PY_OS_URANDOM          (1)
 
diff --git a/ports/windows/mpconfigport.h b/ports/windows/mpconfigport.h
index 55e44c6f5..fabc9072d 100644
--- a/ports/windows/mpconfigport.h
+++ b/ports/windows/mpconfigport.h
@@ -136,7 +136,6 @@
 #define MICROPY_PY_OS_INCLUDEFILE   "ports/unix/modos.c"
 #define MICROPY_PY_OS_ERRNO         (1)
 #define MICROPY_PY_OS_GETENV_PUTENV_UNSETENV (1)
-#define MICROPY_PY_OS_SEP           (1)
 #define MICROPY_PY_OS_STATVFS       (0)
 #define MICROPY_PY_OS_SYSTEM        (1)
 #define MICROPY_PY_OS_URANDOM       (1)