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
RFP with CCAUC Addition
| Types | DnaRegion |
| Roles | Reporter engineered_region |
| Sequences | BBa_K199009_sequence (Version 1) |
Description
This RFP 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 RFP 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 RFP gene, we utilized the restriction enzyme NcoI that cuts at a single site 419-bp into BBa_E1010.Source
The template of the gene comes from BioBrick Part E1010. We designed a forward primer that included ATG, the 5-bp addition CCATC and the first 20 nucleotides on the 5' end of RFP after ATG. This made the edited 5' sequence for our RFP.| Sequence Annotation | Location | Component / Role(s) |
| NcoI Restriction Site 5-bp CCAUC Addition 25 bp addition Double Stop | 424,429 4,8 687,711 681,686 | sequence_feature feature/misc feature/mutation sequence_alteration sequence_feature feature/misc feature/misc sequence_feature |