Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Nicholas Elder
Date created: 2015-11-09 12:00:00
Date modified: 2015-11-10 01:20:24
tCloneTetRed with short stuffer
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_J100238_sequence (Version 1) |
Description
This is the tConeTetRed insert (part BBa_J119386) in pUC-BR plasmid. The insert has had the P2 promoter and GFP gene removed by digestion with BsaI and has had a short stuffer sequence (11bp) ligated in its place. This was used to test the effectiveness of tConeTetRed without the presence of a riboswitch, so the presence of the Tet-A RFP fusion protein is directly controlled by the P5 promoter. This was tested in JM109 E. coli cells and produces measurable fluorescence when grown in the presence of Tetracycline in a range of concentrations from 10 to 50 ug/mL.Notes
When tCloneTetRed is used to test riboswitches, there are approximately 80 bases added between the promoter and ribosome binding site. To test the efficiency of the system without the increased distance between the promoter and RBS, we synthesized this short stuffer sequence to minimize the distance and test the 'natural' activity of the system.Source
The part comes from J119361 tCloneRed and J119140 TetA. The linker sequence was developed from K598018 tetA+GFP fused protein by Qingyang XIAO.The short stuffer sequence is 5' GTGATCCTACA.
| Sequence Annotation | Location | Component / Role(s) |
| P5 promoter Short Stuffer TetA protein half Linker sequence RFP protein half BD18 C Dog RBS | 1,37 41,53 141,1335 1336,1385 1386,2063 53,140 | promoter feature/promoter sequence_feature feature/misc CDS feature/cds feature/misc sequence_feature CDS feature/cds sequence_feature feature/misc |