Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Chris Clifford
Date created: 2012-09-28 11:00:00
Date modified: 2015-05-08 01:13:47
An L-Arabinose Controlled Thiamine Binding - Starch Binding Protein Fusion
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K931009_sequence (Version 1) |
Description
A starch-binding protein gene (CBM21) and a thiamine-binding protein gene (ThiT) were placed in sequence with each other in that ThiT comes before CBM21. CBM21 comes from R. oryzae. It is part of an operon that codes for a two-domain enzyme, one of which binds starch molecules. ThiT comes from E.coli and plays a crucial role in the thiamine transport system by binding the thiamine molecule. It has a Kd of 170 nM when the optimum pH is at 5.0 These two genes were placed after an L. Arabinose promoter sequence (BBa_K206000). This allows the protein to be expressed by adding the L. Arabinose chemical. Two terminator sequences (B0010 B0012) were placed at the end of the construct. The resulting gene construct is 2038 base pairs long. The protein complex from these two genes has a starch-binding domain and a thiamine binding domain which can bind their resulting substrates.Notes
The four genes were placed in sequence of each other with the L. Arabinose promoter coming first, then the thiamine-binding protein gene preceded by the starch-binding protein gene coming next, and then the terminator sequences coming last. The linker sequence between each element is ACTAGA.Source
The four genes come from separate organisms. The L. Arabinose promoter (BBa_K206000) comes from the Registry of Standard Biological Parts and was originally submitted by Group: iGEM09_British_Columbia. Thiamine-binding protein gene comes from the genomic DNA of E. coli and the starch-binding protein gene comes from the genomic DNA of R. oryzae. The two terminator genes (B0010 B0012) come from the Registry of Standard Biological Parts and was originally submitted by Reshma Shetty.| Access | Instance | Definition |
| public public | BBa_K931004 BBa_K931001 | BBa_K931004 BBa_K931001 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K931004 BBa_K931001 | 1,981 988,1785 | BBa_K931004 BBa_K931001 |