Coming up with a

project
Teach the Teachers Workshop
May 16, 2009
Talk to people
Previous iGEM projects
igem.org
New organisms
New parts and tools
for future teams
Most commonly used parts:
B0015 - a terminator
F2620 - an inducible promoter
B0034 - a RBS
R0011 - lac promoter
Plasmid backbones
 BBa_F2620 BBa_F2620
3OC6HSL PoPS Receiver
Mechanism & Function Component Parts
A transcription factor (LuxR) that is active in the presence
of a cell-cell signaling molecule (3OC6HSL) is controlled by
a regulated operator (PLtetO-1). Device input is 3OC6HSL.
Device output is PoPS from a LuxR-regulated operator. If R0040 B0034 C0062 B0015 R0062
used in a cell containing TetR then a second input such as PLtetO-1 RBS luxR Term. Plux,R
aTc can be used to produce a Boolean AND function.

Static Performance* Dynamic Performance*

http://parts.mit.edu/registry/index.php/Part:BBa_F2620
600 8 600 8

GFP synthesis rate (molecules cell!1 s!1)

GFP synthesis rate (molecules cell!1 s!1)

Population Mean
Colony Range 7 + 3OC6HSL 7
500 Hill Equation 500
6 6
400 400
5 5

PoPS cell!1
PoPS cell!1

300 4 300 4

Reuse and
3 3
200 200 GFP synthesis rate (Low Input)
2 GFP synthesis rate (High Input) 2
100 100 Polynomial Fit (High Input)
1 PoPS (High Input) 1

0 0 0 0
0E+00 1E!10 1E!09 1E!08 1E!07 1E!06 1E!05 1E!04 !10 0 10 20 30 40 50
[3OC6HSL] (M) Time (min)

Pmax [3OC6HSL]n Pmax: 6.6 PoPS cell-1 BBa_F2620 Response Time: <1 min

existing parts
Pout = K: 1.5E-09 M 3OC6HSL BBa_T9002 Response Time: 6±1 min
K n + [3OC6HSL]n n: 1.6 Inputs: 0 M (Low), 1E-07 M (High) 3OC6HSL

Input Compatibility* Reliability**

600 8
GFP synthesis rate (molecules cell!1 s!1)

C4HSL
C6HSL 7
500
3OC6HSL
6
C7HSL 92
400
5 74
C8HSL
gs
PoPS cell!1

56
in

3OC8HSL
l

300 38 92
ub

4 1E0

Signaling Devices
GFP 1E1
Do

C10HSL 20
(arb1E2 1E3 74
gs

C12HSL 3 itrar 1E4 56

y un
in

200
its)
l
ub

38
2 Low Input GFP1E0 1E1
Do

(0 M 3OC6HSL) (arb 1E2 1E3 20

100 itrar
1 y un 1E4
its)
High Input
0 0 (1E -7 M 3OC6HSL)
0E+00 1E!10 1E!09 1E!08 1E!07 1E!06 1E!05 1E!04
[AHL] (M) Genetic: >92/>56 culture doublings
Part Compatibility (qualitative) Performance: >92/>56 culture doublings
(low/high input during propagation)
Chassis: MC4100, MG1655, and DH5
Plasmids: pSB3K3 and pSB1A2
Conditions (abridged)
Devices: E0240, E0430 and E0434
Output: PoPS measured via BBa_E0240
Culture: Supplemented M9, 37ºC
Transcriptional Output Demand (low/high input) Plasmid: pSB3K3
Nucleotides: 0 / 6xNt nucleotides cell-1 s-1 Chassis: MG1655
Polymerases: 0 / 1.5E-1xNt RNAP cell-1 *Equipment: PE Victor3 multi-well fluorimeter
(Nt = downstream transcript length) **Equipment: BD FACScan cytometer

Authors: Barry Canton

Ania Labno Registry of Standard Biological Parts License: Public
Updated: March 2008 making life better, one part at a time
Let the students
choose
Help them make smart
choices

• Figure out what’s practical: How many

assembly stages could the team possibly do
over the course of the summer? That sets
an upper limit to the size of the system.
• Design the project so that different
modules can be done in parallel.
• It doesn’t have to be a brand new idea.
Describe your project
on your team wiki
Teach the Teachers Workshop
May 16, 2009