Part:BBa_K766005
T7 promoter-Hig tag-LuxI wihout LVA tag
We build this part to work as the signal sender in our project. T7 promoter is a high-efficient promoter which can bind with T7 polymerase and start the transcription. Besides, we add the lac operator between T7 promoter and CDS.
If you use this part in a lacI-coding plasmid like pet15b, the expression of gene will be repressed. However, IPTG can be used to activate the expression. The expression of this part can be well-controled.
His tag is on the N-terminal, which consists of six histidine amino acid. We can use Ni-column to purify the protein. Besides, you can use Hig-tag antibody to detect the protein in western blot. This design makes the detection of luxI gene become very easy.
On the 5' end of sequence, we add Bgl2 cutting site. On the 5' end of luxI gene, we add Nde1 cuting site. On the 3'end of LuxI gene, we add Xho1 and BamH1 cutting site. All of this design is for the convenience of further use.
This part can be only used in the E.coli strains that can express T7 polymerase like BL21(DE3).
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 67
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1
Illegal BamHI site found at 755
Illegal XhoI site found at 750 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 67
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 67
- 1000COMPATIBLE WITH RFC[1000]
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