Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Leland Taylor
Date created: 2009-06-28 11:00:00
Date modified: 2015-05-08 01:11:18
RFP with CCCUC Addition
| Types | DnaRegion |
| Roles | engineered_region Reporter |
| Sequences | BBa_K199011_sequence (Version 1) |
Description
This RFP reporter gene has a 5-base pair CCACU addition after the start codon and before the rest of the gene. This is used in conjunction with the CCACU tRNA (BioBrick part K199000). This CCACU 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 RFP 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 NcoI which cut at a single site 424 bp into the gene.Source
The template of the gene comes from BioBrick part E1010. 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 RFP gene.| Sequence Annotation | Location | Component / Role(s) |
| CCCUC 5 Base Addition Nco1 restriction site 25 bp addition Double Stop | 4,8 424,429 687,711 681,686 | sequence_feature feature/misc sequence_feature feature/misc feature/misc sequence_feature sequence_feature feature/misc |