Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Olivia Ho-Shing
Date created: 2009-06-28 11:00:00
Date modified: 2015-05-08 01:11:18
Tet Resistance with CCAUC Addition
| Types | DnaRegion |
| Roles | engineered_region Reporter |
| Sequences | BBa_K199010_sequence (Version 1) |
Description
This Tet reporter gene has a 5-bp addition, CCAUC, after the start codon and before the rest of the gene. This is used in conjunction with the CCAUC tRNA (Parts K199007 and K199008). These CCAUC tRNAs suppress the frameshift caused by this 5-bp addition in the reporter.Notes
We positioned the start codon, ATG, before the 5-bp codon and continued with the rest of the Tet gene. This way, the RBS wouldn't translate the reporter gene without reading over the 5-base codon first. In order to ligate our edited 5' sequence to the unchanged downstream portion of the Tet gene, we utilized the restriction enzyme BamHI that cuts at a single site 290-bp into BBa_J31007.Source
The template of the gene comes from BioBrick Part J31007. We designed a forward primer that included ATG, the 5-bp addition CCATC and the first 20 nucleotides on the 5' end of Tet after ATG. This made the edited 5' sequence for our Tet.| Sequence Annotation | Location | Component / Role(s) |
| 5-bp CCAUC Addition BamHI Restriction Site | 4,8 295,300 | sequence_alteration feature/mutation feature/misc sequence_feature |