I've done a couple dozen of these. My guidelines:
When you pregen characters make sure you describe their personality, not their history. In addition, add a section where you say what the character thinks of the other PCs at the table.
Don't be afraid to introduce conflict; so-and-so likes so-and-so, and is in love with so-and-so, but so-and-so is jealous of this. Do this right, and you won't even need an adventure because the roleplaying will take on a life of its own. I usually start with the characters' personalities before I worry about stats and plot.
As for the adventure, limit it to 2 or 3 encounters plus roleplaying -- but have an optional encounter set aside in case they move really fast. No one ever moves really fast. Shoot for a strong beginning and end, and leave the middle a little less linear. Don't be afraid to kill characters in the last encounter if it would make sense.
Expect a four hour game: half an hour to read PCs, three hours to game, half an hour for breaks or chatting.
Encounter Example: in the Spycraft game you almost joined last year at GenCon, my layout looked like this. It was consistently 45 minutes too long, and should have been planned an hour shorter than it is:
1. Briefing. Like prelude, info & questions only. (roleplaying encounter)
2. Drop the PCs in the middle of a car chase on a mountain road. (combat encounter)
3. PCs interrogate survivors. (roleplaying encounter)
4. PCs follow clues, which lead to suspect's apartment. Investigate, probably set off traps. Find clue to where suspect fled. (puzzle & investigation/roleplaying encounter)
5. Fly to Amazon Jungle. Get past drug lord security. (trap/puzzle encounter)
6. Take out two guards. (optional -- combat or stealth encounter)
7. Infiltrate base. (combat or stealth encounter)
8. Kill doctors, blow stuff up and resolve original plot. (combat & roleplaying encounter)
If I was to run this again, I'd make the 2nd encounter (the chase) shorter by eliminating a car of bad guys, and I wouldn't allow the group to dither and stall between encounters 4 and 5. Otherwise, it seemed to be well paced.