Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Michael Florea
Date created: 2014-10-19 11:00:00
Date modified: 2015-05-08 01:09:52
sfGFP fused to CBDcex driven by LacI in pSEVA331-Bb
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1321297_sequence (Version 1) |
Description
This is a LacI-promoter expression construct of super-folder GFP fused N-terminally to CBDcex (a cellulose-binding domain; part BBa_K1321357) expressed in the broad host-range plasmid pSEVA331-Bb (part BBa_K1321300) capable of replication in G. xylinus. BBa_K1321297 is a member of the G.xylinus genetic engineering toolbox.G.xylinus toolbox was designed to ease genetic engineering of cellulose-based biomaterials using the cellulose synthesizing bacterium Gluconacetobacter xylinus (parts Bba_K1321305 and BBa_K1321306). As no registry parts had been tested in G.xylinus, the aim of this toolbox was to determine the parts usable in G.xylinus and characterize them. pSEVA331-Bb is a non-standard broad host range plasmid capable of replication in G.xylinus and E.coli (a shuttle vector) and was selected because the registry's standard plasmid backbone pSB1C3 can not be used for G.xylinus engineering.
NOTE: Because the registry's standard plasmid backbone pSB1C3 is not capable of replication in Gluconacetobacter species, the G.xylinus genetic engineering toolbox is housed mainly in pSEVA331-Bb. pSEVA331-Bb is a non-standard backbone, which therefore can't be quality controlled by and maintained in the registry. However, in order to make the G.xylinus toolbox available for the synthetic biology community, Imperial iGEM 2014 team has made the toolbox freely available upon request, with quality control provided (see Experience). For requests, please contact Imperial iGEM 2014 team.
Notes
BBa_K1321297 was created by restricting BBa_K1321357 and BBa_K1321300 with PstI and XbaI, gel purifying the resulting fragments, ligating using T4 DNA ligase and transforming the ligase products into chemically competent DH10B cellsSource
BBa_K1321357 and BBa_K1321300| Access | Instance | Definition |
| public public | BBa_K1321357 BBa_K1321300 | BBa_K1321357 BBa_K1321300 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K1321357 BBa_K1321300 | 1,1279 1288,4180 | BBa_K1321357 BBa_K1321300 |