Reporter

Part:BBa_K176012

Designed by: Chao Li, Danqian Liu, Hao Jiang   Group: iGEM09_USTC   (2009-06-30)

pCon 0.70(J23101)->RBS+GFP+T

pCon 0.70(J23101)+I13504

Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 705

MEASUREMENT

We measured five constitutive promoter family member ( J23100, J23101, J23103, J23107, J23109, J23115, )strength by ligated each with a 
GFP gene ( I13504) , and then moniter the OD600 and GFP expression.
K176011: J23100 + GFP ( I13504 )
K176012: J23101 + GFP ( I13504)
K176013: K176009( J23107) + GFP ( I13504)
K176014: K176008( J23115) + GFP ( I13504)
K176015: J23109 + GFP ( I13504)
K176016: J23103 + GFP ( I13504)

Model

To see the details of the model, click here please.

data

J23101+GFP

COLONY NUMBER T(min) OD600 FLU(arbitrary) ST.FLU(ustc_st.1) FLU/OD600(arbitrary) ST.FLU/OD600(ustc_st.1)
0 0.117666667 142.91 571.2515489 1214.532578 4854.829028
30 0.119666667 152.6266667 610.0918042 1275.431755 5098.260203
60 0.143666667 181.65 726.1062478 1264.385151 5054.103813
90 0.174666667 206.08 823.7598433 1179.847328 4716.182309
120 0.198 220.6666667 882.0668612 1114.478114 4454.883137
150 0.245666667 242.8866667 970.8864639 988.6838535 3952.048021
180 0.392 276.32 1104.52892 704.8979592 2817.675817
0 0.083 115.0533333 459.9006009 1386.184739 5540.971095
30 0.098333333 126.3533333 505.0698858 1284.949153 5136.303924
60 0.112 144.0466667 575.795126 1286.130952 5141.027911
90 0.129666667 188.7433333 754.4603003 1455.604113 5818.4599
120 0.14 174.4166667 697.1925757 1245.833333 4979.946969
150 0.169333333 184.28 736.619099 1088.267717 4350.112789
180 0.251333333 232.6733333 930.0608919 925.7559682 3700.507528
For the details of the definition of ST.FLU and the unit ustc_st1, please click here. 

plot

Promoters.jpg

protocol

1. Streak a plate of the strain which contain one of the parts listed in pSB1A3 .

2. Inoculate two 3ml cultures of supplemented M9 Medium and antibiotic( Ampicillin 0.1mg/ml) with single colony from the plate.

3. Cultures were grown in test tubes(BIO BASIC INC.12ml Polypropylene Round-bottom Culture Tubes With Graduations And Dual Cap Cat.No:TD444) for 16hrs at 37℃ with shaking at 200rpm.

4. Cultures were diluted 1:100 into 3ml fresh medium and grown for 3hrs.

5. Measure the fluorescence(SHIMDZU SPECTROFLUOROPHOTOMETER RF-5301PC, 250ul quartz cell path length 10mm,501 nm excitation,514 nm emission,1.5nm slit width) and absorbance (HITACHI UV-VIS spectrophotometer U-2810 ,200ul quartz cell,path length 10mm,600nm,1.5 nm slit width) every 30 minutes in the next 4hrs.


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Categories
Parameters
n/apCon 0.70(J23101)->RBS+GFP+T