Part:BBa_K613013
TetR V36F mutant
TetR with a point mutation V36F.
In vitro characterization
PO | A | T | C | G |
1 | 0.164072 | 0.519334 | 0 | 0.22966 |
2 | 0.642569 | 0.164072 | 0.188965 | 0.943314 |
3 | 1.04481 | 0.558693 | 0.164072 | 0.848264 |
4 | 0.959646 | 0.164072 | 2.17536 | 0.718959 |
5 | 0.164072 | 1.19894 | 1.79469 | 1.56685 |
6 | 1.46463 | 0.164072 | 1.74384 | 1.54845 |
7 | 0.966397 | 1.07557 | 0.164072 | 1.72883 |
8 | 0.164072 | 0.504312 | 0.755195 | 0.148902 |
9 | 0.0876645 | 0.164072 | 0 | 0.0156484 |
10 | 0.289722 | 0.164072 | 0.00707563 | 0.843474 |
11 | 1.77568 | 1.3447 | 2.34804 | 0.164072 |
12 | 0.164072 | 1.72354 | 1.62782 | 1.49176 |
13 | 0.877518 | 0.164072 | 1.30879 | 1.9052 |
14 | 0.164072 | 0.544642 | 0.3387 | 1.7537 |
15 | 0.540091 | 0.8821 | 1.0861 | 0.164072 |
16 | 0 | 0.280358 | 0.442769 | 0.164072 |
17 | 0.31238 | 0 | 0.164072 | 0 |
ΔΔG° values are indicated in kcal/mol
In vivo characterization
This TetR mutant was characterized in vivo by putting it into pSB3K1 under a constitutive promoter (J23116). This plasmid was cotransformed with J61002 harbouring RFP under pTet promoter (B0040) in DH5alpha cells. Cells were grown in a medium containing Kanamycin & Amplicillin plus different concentrations of ATC, ranging from 0 to 2000 ng/mL. OD600 absorbence and RFP fluorescence were measured every 10 minutes during 12 hours on a platereader machine.
Induction curves
Fluorescence measurements (RFUs) were normalized by OD600 values.
Dose-response curve
Fluorescence measurements (RFUs) were normalized by OD600 values. For each ATC concentration, we estimated the steady-state fluorescence expression by averaging the measurements over the last hour.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
None |