Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Fiona Tsai
Date created: 2016-10-09 11:00:00
Date modified: 2016-10-19 09:50:48
Strong promoter + Strong RBS + GSR + 10x Histidine tag + Double terminator
| Types | DnaRegion |
| Roles | Composite engineered_region |
| Sequences | BBa_K1898250_sequence (Version 1) |
Description
BBa_K880005 is used to provide a strong promoter and strong rbs to maximize protein output. The ORF includes GSR, or glutathione reductase that catalyses the conversion of glutathione disulfide to glutathione, and 10x Histidine tag for protein purification. BBa_B0015 consists of a double terminator to end transcription.Notes
We did five silent point mutations on gsr to remove the two Pst1 and three EcoR1 internal cutting sites. The sequence was then sent to Missiontech for mutagenesis. Primers were designed to remove the stop codon from GSR and were synthesized by Tri-I. Our assembly method BioBrick RFC[10] shifted the open reading frame and created a stop codon in between GSR and 10x histidine tag. The sequence was sent to Missiontech to add two base pairs in between the two genes.Source
the cDNA of GSR was ordered from OriGene. The promoter and rbs was from iGEM distribution kit (BBa_K880005) and His-Term was from an intermediate part we constructed (BBa_K1898500).| Access | Instance | Definition |
| public public public | BBa_K880005 BBa_K1898200 BBa_S05336 | BBa_K880005 BBa_K1898200 BBa_S05336 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K880005 BBa_K1898200 BBa_S05336 | 1,55 62,1627 1636,1808 | BBa_K880005 BBa_K1898200 BBa_S05336 |