BBa_J100066

BBa_J100066 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_J100066
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Rebecca Evans
Date created: 2012-06-11 11:00:00
Date modified: 2015-08-31 04:08:22

Synthetic Riboswitch



Types
DnaRegion

Roles
promoter

Regulatory

Sequences BBa_J100066_sequence (Version 1)

Description

This part is a synthetic riboswitch that can be used in E. coli. In the presence of theophylline, the expression of the gene of interest is induced, but in the absence of theophylline, very little transcription takes place. This part contains a modified T5 promoter, which, in the absence of cymR, acts as a strong constitutive promoter. The ribosomal binding site is contained in the riboswitch.

Because the riboswitch must be right beside the gene of interest (the start codon is folded in the riboswitch), we added a BsaI site. This allows the riboswitch to be attached to the gene of interest using Golden Gate Assembly (as long as the first nucleotide after the start codon of the gene of interest is a C, we designed the part to use with superfolder GFP).

This riboswitch is modified from riboswitch E as described in the paper by Topp et al.

References: Shana Topp, Colleen M. K. Reynoso, Jessica C. Seeliger, Ian S. Goldlust, Shawn K. Desai, Doroth??e Murat, Aimee Shen, Aaron W. Puri, Arash Komeili, Carolyn R. Bertozzi, June R. Scott, and Justin P. Gallivan. Synthetic Riboswitches That Induce Gene Expression in Diverse Bacterial Species. Applied and Environmental Microbiology, 76:23, 7881-7884. December 2010.

Notes

None

Source

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2988590/?tool=pubmed

Sequence Annotation Location Component / Role(s)
BBa prefix
T5 promoter
lac operator
scar
constant sequence
KpnI site
Riboswitch
Start
RBS
GGA suffix ATGC sticky word
BBa suffix
1,22
23,75
76,96
97,102
104,118
119,124
125,187
185,187
168,174
185,195
196,216
feature/BioBrick engineered_region
feature/promoter promoter
feature/operator operator
sequence_feature feature/misc
sequence_feature feature/misc
sequence_feature feature/misc
sequence_feature feature/misc
feature/start start_codon
feature/rbs ribosome_entry_site
feature/misc sequence_feature
feature/BioBrick engineered_region
igem#sampleStatus
Not in stock
igem#status
Unavailable
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_J100066/1