Part:BBa_K3447112
light-off induced system
This part can turn off the expression of the sfGFP gene when blue light irradiation.
YF1 is the kinase for FixJ in the blue light system. Without blue light irradiation, YF1 phosphorylates FixJ, activating the downstream expression after promoter PFixK2. Once the blue light is on, the FixJ cannot be phosphorylated, shutting down the downstream gene expression.
Characterization
In the blue light-off system, YF1 switched to turn on in the dark and phosphorylate FixJ, which binds to the PFixK2 to initiate expression of the downstream sfGFP. When irradiated with blue light, the phosphorylation of YF1 was repressed, thereby inhibiting the downstream sfGFP (Fig. 1A & B).
To verify the construction and function of blue light-off system, two groups were set to proof the strong inhibition of the blue light on our bacteria. Moreover, compared with the construct without blue light, our blue light-off system would constantly inhibit the fluorescent expression with blue light induced. And since the construct shows little difference in fluorescent expression with the mock when blue light on, we indicated our system with a low leak (Fig. 2 and 3).
Design
Design Notes
We added some synonymous mutations to avoid part rules.
Source
We found this sequence data in GenBank.
References
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 3041
Illegal NheI site found at 3064 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1882
Illegal XhoI site found at 2590
Illegal XhoI site found at 3125 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 587
Illegal NgoMIV site found at 659
Illegal NgoMIV site found at 749
Illegal NgoMIV site found at 767
Illegal NgoMIV site found at 1259
Illegal NgoMIV site found at 1552
Illegal NgoMIV site found at 1646
Illegal AgeI site found at 301
Illegal AgeI site found at 1427 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1316
Illegal BsaI.rc site found at 200
None |