"
Team:Uppsala-SwedenWeek6
From 2010.igem.org
Week-6
Design of the plasmids and biobricks
We decided to design 3 band-detect constructs. Each one with the same set up of genes but differentiating by the promotors sensitivity of activators or repressors and the fluorescent proteins are different between the constructs as well.
Plasmid representation:
We used the Arhenius software to get a pictorial design of our plasmids. Please find the images of the different plasmids given below. These images were very helpful in the flowchart design stage of our project. Moreover having a pictorial representation of the project made it easier to explain it to others.
Flowchart for Construction of the plasmids:
|
Biobricks details |
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|
Part BBa_ |
Name |
PCR length |
Length (bp) |
Plasmid Length |
Total Length=B2+C2 |
Plasmid |
Plasmid number |
Plate |
Well |
Antibiotics |
Sequencing |
Quality (Gel, Q, P) |
Function |
|
|
F 2621 |
F2 |
1396 |
1158 |
2079 |
3237 |
PSB1A2 |
All |
2 |
21F |
Amp |
Confirmed |
Bad, OK, OK |
Sensor |
|
|
F 1610 |
F1 |
1114 |
798 |
3189 |
3987 |
pSB1Ak3 |
AHL Relay |
2 |
24G |
Amp & Km |
Confirmed |
OK, OK, OK |
AHL generator |
|
|
I 746350 |
F17 |
475 |
237 |
2079 |
2316 |
pSB1A2 |
1 & 4 |
2 |
12C |
Amp |
Partially |
OK, OK, OK |
Activator in sensitivity tuner |
|
|
I 746352 |
F4 |
502 |
264 |
2079 |
2343 |
pSB1A2 |
2,3,5 & 6 |
2 |
12G |
Amp |
Partially |
OK, OK, OK |
Activator in sensitivity tuner |
|
|
B0015 |
F5 |
445 |
129 |
3189 |
3318 |
pSB1AK3 |
All |
1 |
23L |
Amp & Km |
Confirmed |
OK, OK, OK |
Terminator |
|
|
I746360 |
F6 |
329 |
91 |
2079 |
2170 |
pSB1A2 |
1 & 4 |
2 |
12I |
Amp |
Confirmed |
OK, OK, OK |
Lower level promotor |
|
|
I746364 |
F7 |
331 |
93 |
2079 |
2172 |
pSB1A2 |
2 & 5 |
2 |
12O |
Amp |
Confirmed |
OK, OK, OK |
Lower level promotor |
|
|
I746365 |
F8 |
330 |
92 |
2079 |
2171 |
pSB1A2 |
3 & 6 |
2 |
14A |
Amp |
Inconsistent |
OK, OK, OK |
Lower level promotor |
|
|
Q04510 |
F9 |
1303 |
987 |
4425 |
5412 |
pSB2K3 |
All |
1 |
18B |
Km |
Confirmed |
OK, Low, OK |
Inverter |
|
|
P0412 |
F10 |
1546 |
1308 |
2079 |
3387 |
pSB1A2 |
All |
3 |
6P |
Amp |
Confirmed |
OK, OK, OK |
Lower level repressor |
|
|
I746361 |
F11 |
330 |
92 |
2079 |
2171 |
pSB1A2 |
1,2,4 & 5 |
2 |
12K |
Amp |
Confirmed |
OK, OK, OK |
Higher level promotor |
|
|
R0010 |
F12 |
438 |
200 |
2079 |
2279 |
pSB1A2 |
4,5 & 6 |
1 |
1D |
Amp |
Confirmed |
OK, OK, OK |
Regulatory promotor |
|
|
I0460 |
F16 |
1285 |
969 |
3189 |
4158 |
pSB1AK3 |
4,5 & 6 |
2 |
24I |
Amp & Km |
Partially |
OK, OK, OK |
AiiA generator |
|
|
I13504 |
F13 |
1113 |
875 |
2079 |
2954 |
pSB1A2 |
1 (GFP) |
1 |
22I |
Amp |
Confirmed |
OK, High, OK |
GFP Generator |
|
|
I13507 |
F14 |
1099 |
861 |
2079 |
2940 |
pSB1A2 |
2 (RFP) |
1 |
22O |
Amp |
Confirmed |
OK, High, OK |
RFP Generator |
|
|
E0430 |
F15 |
1116 |
878 |
2079 |
2957 |
pSB1A2 |
3 (YFP) |
1 |
8I |
Amp |
Confirmed |
OK, High, OK |
YFP Generator |
|
|
B0034 |
F3 |
250 |
12 |
2079 |
2091 |
pSB1A2 |
4,5 & 6 |
1 |
2M |
Amp |
Confirmed |
OK, High, OK |
RBS |
|
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|
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|
F2621-F1610 |
G1 |
2262 |
1956 |
2072 |
4028 |
pSB1C3 |
|
|
|
|
|
|
|
|
|
R0010-F1610 |
G2 |
1312 |
1006 |
2072 |
3078 |
pSB1C3 |
|
|
|
|
|
|
|
|
|
F2621-I746350 |
G3 |
1718 |
1403 |
2206 |
3609 |
pSB1K3 |
|
|
|
|
|
|
|
|
|
F2621-I746352 |
G4 |
1745 |
1430 |
2206 |
3636 |
pSB1K3 |
|
|
|
|
|
|
|
|
|
Q04510-P0412 |
G5 |
2609 |
2303 |
2072 |
4375 |
pSB1C3 |
|
|
|
|
|
|
|
|
|
P0412-R0010 |
G6 |
1822 |
1516 |
2072 |
3588 |
pSB1C3 |
|
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F2621-I746350-B0015 |
H1 |
1846 |
1540 |
2072 |
3612 |
pSB1C3 |
|
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F2621-I746352-B0015 |
H2 |
1873 |
1567 |
2072 |
3639 |
pSB1C3 |
|
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I746360-Q04510-P0412 |
H3 |
2717 |
2402 |
2206 |
4608 |
pSB1K3 |
|
|
|
|
|
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|
|
I746361-Q04510-P0412 |
H4 |
2718 |
2403 |
2206 |
4609 |
pSB1K3 |
|
|
|
|
|
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|
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I746364-Q04510-P0412 |
H5 |
2719 |
2404 |
2206 |
4610 |
pSB1K3 |
|
|
|
|
|
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|
|
I746361-P0412-R0010 |
H6 |
1931 |
1616 |
2206 |
3822 |
pSB1K3 |
|
|
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I746364-P0412-R0010 |
H7 |
1932 |
1617 |
2206 |
3823 |
pSB1K3 |
|
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|
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I746365-P0412-R0010 |
H8 |
1931 |
1616 |
2206 |
3822 |
pSB1K3 |
|
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F2621-I746350-B0015-I746360-Q04510-P0412 |
I1 |
4265 |
3950 |
2157 |
6107 |
pSB1A3 |
|
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|
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F2621-I746352-B0015-I746361-Q04510-P0412 |
I2 |
4293 |
3978 |
2157 |
6135 |
pSB1A3 |
|
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F2621-I746352-B0015-I746364-Q04510-P0412 |
I3 |
4294 |
3979 |
2157 |
6136 |
pSB1A3 |
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F2621-I746350-B0015-I746360-Q04510-P0412-I746361-P0412-R0010 |
J1 |
5890 |
5574 |
2463 |
8037 |
pSB1T3 |
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F2621-I746352-B0015-I746361-Q04510-P0412-I746364-P0412-R0010 |
J2 |
5919 |
5603 |
2463 |
8066 |
pSB1T3 |
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F2621-I746352-B0015-I746364-Q04510-P0412-I746365-P0412-R0010 |
J3 |
5919 |
5603 |
2463 |
8066 |
pSB1T3 |
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F2621-I746350-B0015-I746360-Q04510-P0412-I746361-P0412-R0010-I13504 |
K1 [http://partsregistry.org/Part:BBa_K379000 BBa_K379000] |
6691 |
6385 |
2072 |
8457 |
pSB1C3 |
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F2621-I746352-B0015-I746361-Q04510-P0412-I746364-P0412-R0010-I13507 |
K2 [http://partsregistry.org/Part:BBa_K379001 BBa_K379001] |
7493 |
7187 |
2072 |
9259 |
pSB1C3 |
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F2621-I746352-B0015-I746364-Q04510-P0412-I746365-P0412-R0010-E0430 |
K3 [http://partsregistry.org/Part:BBa_K379002 BBa_K379002] |
6723 |
6417 |
2072 |
8489 |
pSB1C3 |
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F2621-I746350-B0015-I746360-Q04510-P0412-I746361-P0412-R0010-I0460 |
K4 |
6785 |
6479 |
2072 |
8551 |
pSB1C3 |
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F2621-I746352-B0015-I746361-Q04510-P0412-I746364-P0412-R0010-I0460 |
K5 |
7601 |
7295 |
2072 |
9367 |
pSB1C3 |
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F2621-I746352-B0015-I746364-Q04510-P0412-I746365-P0412-R0010-I0460 |
K6 |
6814 |
6508 |
2072 |
8580 |
pSB1C3 |
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Characterization
Coming soon
