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sample code, reproducible

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import tkinter as tk
import win32gui
import win32con

root = tk.Tk()
root.geometry('500x500+%s+%s'%(200, 130))

root.overrideredirect(1)
root.config(bg='#fff')
root.attributes('-topmost', True)
root.attributes('-alpha', 0.25)

hwnd = root.winfo_id()

lExStyle = win32gui.GetWindowLong(hwnd, win32con.GWL_EXSTYLE)
lExStyle |=  win32con.WS_EX_TRANSPARENT | win32con.WS_EX_LAYERED
win32gui.SetWindowLong(hwnd, win32con.GWL_EXSTYLE , lExStyle )

root.mainloop()