Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Shamita Punjabi
Date created: 2009-06-25 11:00:00
Date modified: 2015-05-08 01:11:18
Tet Resistance with CCACU Addition
| Types | DnaRegion |
| Roles | Reporter engineered_region |
| Sequences | BBa_K199006_sequence (Version 1) |
Description
This Tet reporter gene has a 5-base pair CUAGU addition after the start codon and before the rest of the gene. This is used in conjunction with the CUAGU tRNA K199002).This CUAGU tRNA suppresses the frameshift caused by this 5-base pair addition in the reporter.
Notes
We positioned the ATG before the 5-base codon and continued with the rest of the Tet resistance gene. This way, the RBS wouldn't read the reporter gene without reading over the 5-base codon first. In order to ligate our 5-base pair addition at the desired position in the reporter gene, we utilized the restriction enzyme BamHI which cut at a single site 290 bp into the gene.Source
The template of the gene comes from BioBrick part J31007. We designed a forward primer that included the first 20 nucleotides on the 5' end, and a reverse primer that included the last 20 nucleotides on the 3' end of the Tet resistance gene.| Sequence Annotation | Location | Component / Role(s) |
| 5 base pair addition BamHI restriction site | 4,8 295,300 | sequence_feature feature/misc sequence_feature feature/misc |