Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Lai Hei Ming
Date created: 2014-11-02 12:00:00
Date modified: 2015-05-08 01:10:23
Gene cluster for Expressing a purifiable Eut Bacterial Microcompartment
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1428001_sequence (Version 1) |
Description
The following features of the gene cluster are listed from upstream to downstream: A T7 promoter, a T7 RBS, His6-tagged EutS, a Plac promoter, an RBS (B0034), EutM, an RBS (B0034), EutN, a Plac promoter, an RBS (B0034), EutL, an RBS (B0034), EutK.The whole gene cluster encodes five proteins, EutS, EutM, EutN, EutL, EutK, arranged in a specific order such that when induced with 0.2mM IPTG or allowed for leaky expression at 30C overnight, these 5 gene products will be expressed in a specific molar ratio so that they can assemble into the intact organelle of the bacterial microcompartment (BMC) of the Ethanolamine-utilization type (Eut). It was derived from BBa_K311004 and the pET28a vector in 2 cloning steps.
If the whole part is put into a bacterial, one can readily express out a whole organelle in the form of BMC for compartmentalization, which, when combined with our second Biobrick BBa_K1428000, can target any coding Biobrick inside the BMC for both in vivo and in vitro applications, for example, metabolic engineering to avoid toxic intermediate damage to the cell, bioremediation for pollutant collection, drug delivery systems by storing nucleic acids, peptides and proteins for replacement therapy, or even virology model studies as the whole thing resembles a viral capsid.
Notes
Careful considerations about the linker peptide between the His6-tag and EutS must be considered to avoid (1) out-frame fusion and (2) linker peptides which are hydrophobic and inflexible.Source
BBa_K311004, pET28a| Sequence Annotation | Location | Component / Role(s) |
| T7 Promoter lac operator T7 RBS Start codon His6-tag Thrombin site EutS CAP binding site Modified Plac Promoter RBS EutM EutN CAP binding site Modified Plac Promoter RBS EutL EutK | 1,19 20,44 64,80 88,90 100,117 127,144 148,483 599,620 635,665 689,696 703,993 1106,1393 1509,1530 1545,1575 1599,1606 1613,2272 2285,2779 | feature/promoter promoter sequence_feature feature/misc ribosome_entry_site feature/rbs feature/start start_codon CDS feature/cds feature/cds CDS CDS feature/cds sequence_feature feature/misc feature/promoter promoter ribosome_entry_site feature/rbs feature/cds CDS CDS feature/cds feature/misc sequence_feature promoter feature/promoter sequence_feature feature/misc CDS feature/cds feature/cds CDS |