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Not sure if this is worth being a list, but if we make one, here's my rough draft for the top 20.

Criteria: Influence, Acclaim, Popularity
1. The Tramp (Charles Chaplin) - City Lights (1931)
2. James Bond (Sean Connery) - Dr. No (1962)
3. Norman Bates (Anthony Perkins) - Psycho (1960)
4. Darth Vader (David Prowse / James Earl Jones - voice) - The Empire Strikes Back (1980)
5. Mickey Mouse (Walt Disney) - Steamboat Willie (1927)
6. Bugs Bunny (Mel Blanc) - Porky's Hare Hunt (1938)
7. Vito Corleone (Marlon Brando) - The Godfather (1972)
8. Indiana Jones (Harrison Ford) - Raiders of the Lost Ark (1981)
9. Scarlett O'Hara (Vivien Leigh) - Gone with the Wind (1939)
10. Hannibal Lecter (Anthony Hopkins) - The Silence of the Lambs (1991)
11. Charles Foster Kane (Orson Welles) - CItizen Kane (1941)
12. Michael Corleone (Al Pacino) - The Godfather (1972)
13. King Kong (n/a) - King Kong (1933)
14. Godzilla (n/a) - Godzilla (1954)
15. Dorothy Gale (Judy Garland) - The Wizard of Oz (1939)
16. Atticus Finch (Gregory Peck) - To Kill a Mockingbird (1962)
17. The Wicked Witch of the West (Margaret Hamilton) - The Wizard of Oz (1939)
18. The Man with No Name (Clint Eastwood) - A Fistful of Dollars (1964)
19. Frankenstein's Monster (Boris Karloff) - Frankenstein (1931)
20. Hal-9000 (Douglas Rain) - 2001: A Space Odyssey (1968)
21. Hans Beckert (Peter Lorre) - M (1931)
22. The Queen (Lucille Laverne) - Snow White and the Seven Dwarfs (1937)
23. Rick Blaine (Humphrey Bogart) - Casablanca (1942)
24. Marshal Will Kane (Gary Cooper) - High Noon (1952)
25. Tony Montana (Al Pacino) - Scarface (1983)
26. Joker (Heath Ledger) - The Dark Knight (2008)
27. Forrest Gump (Tom Hanks) - Forrest Gump (1994)
28. Luke Skywalker (Mark Hamill) - Star Wars (1977)
29. George Bailey (James Stewart) - It's a Wonderful Life (1946)
30. Han Solo (Harrison Ford) - Star Wars (1977)
31. Batman (Christian Bale) - Batman Begins (2005)
32. Popeye the Sailor (Billy Costello) - Popeye the Sailor (1933)
33. Rocky Balboa (Sylvester Stallone) - Rocky (1976)
34. The Moon (Georges Melies) - A Trip to the Moon (1902)
35. T.E. Lawrence (Peter O'Toole) - Lawrence of Arabia (1962)
36. T-800 (Arnold Schwarzenegger) - The Terminator (1984)
37. Joan of Arc (Renee Jeanne Falconetti) - The Passion of Joan of Arc (1928)
38. Travis Bickle (Robert De Niro) - Taxi Driver (1976)
39. Totoro (Hitoshi Takaga) - My Neighbor Totoro (1988)
40. Ellen Ripley (Sigourney Weaver) - Alien (1979)

I mean based on the criteria...Mickey Mouse could arguably be #1, right?

Maybe, but a pretty large part of the Mouse's popularity is from television and Disney's overall brand image, though most of his influence would definitely go to the films. I might move him up to #5.

Technically it's Frankenstein's Monster, Man.

So if we are to roughly split this in heroes/villains category:

Heroes:

1. The Tramp (Charles Chaplin) - City Lights (1931)
2. James Bond (Sean Connery) - Dr. No (1962)
3. Indiana Jones (Harrison Ford) - Raiders of the Lost Ark (1981)
4. Dorothy Gale (Judy Garland) - The Wizard of Oz (1939)
5. Atticus Finch (Gregory Peck) - To Kill a Mockingbird (1962)
6. The Man with No Name (Clint Eastwood) - A Fistful of Dollars (1964)
7. Rick Blaine (Humphrey Bogart) - Casablanca (1942)
8. Marshal Will Kane (Gary Cooper) - High Noon (1952)
9. Forrest Gump (Tom Hanks) - Forrest Gump (1994)
10. Luke Skywalker (Mark Hamill) - Star Wars (1977)
11. George Bailey (James Stewart) - It's a Wonderful Life (1946)
12. Han Solo (Harrison Ford) - Star Wars (1977)
13. Batman (Christian Bale) - Batman Begins (2005)
14. Popeye the Sailor (Billy Costello) - Popeye the Sailor (1933)
15. Rocky Balboa (Sylvester Stallone) - Rocky (1976)
16. T.E. Lawrence (Peter O'Toole) - Lawrence of Arabia (1962)
17. Joan of Arc (Renee Jeanne Falconetti) - The Passion of Joan of Arc (1928)
18. Ellen Ripley (Sigourney Weaver) - Alien (1979)

Villains:

1. Norman Bates (Anthony Perkins) - Psycho (1960)
2. Darth Vader (David Prowse / James Earl Jones - voice) - The Empire Strikes Back (1980)
3. Vito Corleone (Marlon Brando) - The Godfather (1972)
4. Hannibal Lecter (Anthony Hopkins) - The Silence of the Lambs (1991)
5. Charles Foster Kane (Orson Welles) - CItizen Kane (1941)
6. Michael Corleone (Al Pacino) - The Godfather (1972)
7. The Wicked Witch of the West (Margaret Hamilton) - The Wizard of Oz (1939)
8. HAL-9000 (Douglas Rain) - 2001: A Space Odyssey (1968)
9. Hans Beckert (Peter Lorre) - M (1931)
10. The Queen (Lucille Laverne) - Snow White and the Seven Dwarfs (1937)
11. Tony Montana (Al Pacino) - Scarface (1983)
12. Joker (Heath Ledger) - The Dark Knight (2008)
13. T-800 (Arnold Schwarzenegger) - The Terminator (1984)
14. Travis Bickle (Robert De Niro) - Taxi Driver (1976)

Grey area characters:

1. Mickey Mouse (Walt Disney) - Steamboat Willie (1927)
2. Bugs Bunny (Mel Blanc) - Porky's Hare Hunt (1938)
3. Scarlett O'Hara (Vivien Leigh) - Gone with the Wind (1939)
4. King Kong (n/a) - King Kong (1933)
5. Godzilla (n/a) - Godzilla (1954)
6. Frankenstein's Monster (Boris Karloff) - Frankenstein (1931)
7. The Moon (Georges Melies) - A Trip to the Moon (1902)
8. Totoro (Hitoshi Takaga) - My Neighbor Totoro (1988)

I added Bugs Bunny and Dorothy Gale to the top 20.

Expanded it to 40.

Thinking about characters from the 2000's - I would consider maybe Captain Jack Sparrow or some of the characters from LOTR over Bale's Batman, just a thought. Ledger's Joker seems appropriately placed though.

I'm counting every iteration of Batman, not just Bale's, since that's clearly what the hero and villain lists were doing as well.

Yes but currently on the OG Villains list have separate entries for Ledger's Joker + Jack Nicholson's Joker (and I'm assuming once we update the list we will include Phoenix as well). So is there no room to include Keaton's Batman as well?

*probably no room for Jared Leto's "interpretation" of the Joker...sadly

I'm not sure I agree with separating them, since the list is ranking characters and not portrayals, but others may feel differently.

Cool list idea! I agree with Tim that it should be Frankenstein's monster.

Corrected.