BBa_K199012

BBa_K199012 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K199012
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

Tet Resistance with CCCUC Addition



Types
DnaRegion

Roles
Reporter

engineered_region

Sequences BBa_K199012_sequence (Version 1)

Description

This Tet resistance reporter gene has a 5-base pair CCCUC addition after the start codon and before the rest of the gene. This is used in conjunction with the CCCUC tRNA (BioBrick part K199001). This CCCUC 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)
CCCUC 5-base pair addition
BamHI Restriction Site
4,8
295,300
sequence_feature feature/misc
sequence_feature feature/misc
igem#experience
Works
 
igem#sampleStatus
It's complicated
igem#status
Planning
 
synbiohub#ownedBy
user/myers
 
synbiohub#ownedBy
user/james
 
synbiohub#topLevel
BBa_K199012/1