Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Eric Holmes
Date created: 2014-08-13 11:00:00
Date modified: 2016-12-03 10:52:20
pT7 + RBS + GST (glutathione-S-transferase)-CRS5 (metallothionein) + Ter
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1460001_sequence (Version 1) |
Description
This part serves as a functional metallothionein for heavy metal binding and conferred resistance to heavy metals when combined with a strain expressing the bacteriophage T7 polymerase. CRS5 gene is the metallothionein that binds to heavy metals and it is fused to GST for stability.Notes
Flanking the T7 promoter are two KpnI cut sites with both the T7 promoter and a SacI cut site between them. These cut sites are present for easy removal of the T7 promoter for modulation of parts with other promoters. The SacI cut site can be used as a marker for successful removal of the T7 promoter from the part. When digested with SacI, part BBa_K1460001 in pSB1C3 will yield bands of size 2166bp and 931bp. If the T7 promoter is successfully removed, when digested with SacI the plasmid will yield the linearized length of the pSB1C3+BBa_K1460001Source
T7 promoter is part BBa_I712074. RBS is from part BBa_J61101. Glutathione-S-transferase is from S. japonicum. CRS5 is a metallothionein from S. cerevisiae. Terminator is part BBa_B1006. This sequence was synthesized by GenScript in the vector pUC57 and was then transferred using traditional cloning into pSB1C3.| Sequence Annotation | Location | Component / Role(s) |
| T7 Promoter BBa_J61101 GST Linker Sequence CRS5 BBa_B1006 | 13,59 72,85 92,745 746,778 779,988 989,1027 | feature/promoter promoter ribosome_entry_site feature/rbs CDS feature/cds feature/misc sequence_feature CDS feature/cds sequence_feature feature/misc |