Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Yu Zhao
Date created: 2012-09-23 11:00:00
Date modified: 2015-05-08 01:13:14
T7 promoter-Hig tag-LuxI wihout LVA tag
| Types | DnaRegion |
| Roles | Generator engineered_region |
| Sequences | BBa_K766005_sequence (Version 1) |
Description
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).
Notes
We want to build a signal sender which is easy detected and controled.Source
We clone this gene from our lab's plamid which is commonly used| Sequence Annotation | Location | Component / Role(s) |
| T7 promoter Lac operator Poly his tag LuxI without LVA tag T7 terminator | 20,37 39,65 120,137 177,746 819,866 | feature/promoter promoter feature/operator operator tag feature/tag CDS feature/cds feature/stop stop_codon |