Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Alejandro Gutierrez
Date created: 2015-09-09 11:00:00
Date modified: 2015-09-11 02:43:05
Device to demethylate 3-methylxanthine
| Types | DnaRegion |
| Roles | Device engineered_region |
| Sequences | BBa_K1627001_sequence (Version 1) |
Description
This part contains genes that allow for the demethylation of 3-methylxanthine. The part contains a synthetic operon consisting of ndmB, ndmD from Pseudomonas putida CBB5, and gst9 from Janthinobacterium sp. Marseille under the control of a constitutive promoter, J23110.This part can be used in a ∆guaB strain of E. coli to measure the amount of 3-methylxanthine in a solution. By growing the ∆guaB strain with this device in a minimal media with different known concentrations of 3-methylxanthine, a growth curve can be made that correlates culture growth (OD600) with concentration of 3-methylxanthine. Then if the strain is grown in a minimal media with an unknown concentration of 3-methylxanthine, the growth can be compared to the growth curve to calculate the initial concentration of 3-methylxanthine in the media.
Notes
These genes were amplified from the pDCAF3 plasmid (BBa_K734000). An EcoRI restriction site was removed from the ndmB gene during the cloning process. The genes were cloned into the pSB1C3 backbone using BioBrick Standard Assembly.Source
The ndmB and ndmD genes were isolated from Pseudomonas putida CBB5. gst9 was isolated from Janthinobacterium sp. Marseille.| Sequence Annotation | Location | Component / Role(s) |
| J23110 K082001 ndmB B0032 ndmD B0034 gst9 | 1,35 44,59 65,1132 1141,1153 1160,2926 2971,2982 2989,3663 | feature/promoter promoter ribosome_entry_site feature/rbs feature/cds CDS ribosome_entry_site feature/rbs feature/cds CDS ribosome_entry_site feature/rbs CDS feature/cds |