Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Alejandro Gutierrez
Date created: 2015-09-09 11:00:00
Date modified: 2015-09-17 07:30:00
Device to demethylate paraxanthine
| Types | DnaRegion |
| Roles | engineered_region Device |
| Sequences | BBa_K1627004_sequence (Version 1) |
Description
This part contains genes that allow for the demethylation of paraxanthine. The part contains a synthetic operon consisting of ndmA, ndmC, and 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 paraxanthine in a solution. By growing the ∆guaB strain with this device in a minimal media with different known concentrations of paraxanthine, a growth curve can be made that correlates culture growth with concentration of paraxanthine. Then if the strain is grown in a minimal media with an unknown concentration of paraxanthine, the growth can be compared to the growth curve to calculate the initial concentration of paraxanthine in the media.
Notes
These genes were amplified from the pDCAF3 plasmid (BBa_K734000). A PstI restriction site was removed from the ndmA gene during the cloning process. The genes were cloned into the pSB1C3 backbone using BioBrick Standard Assembly.Source
The ndmA, ndmC, and ndmD genes were isolated from Pseudomonas putida CBB5. gst9 was isolated from Janthinobacterium sp. Marseille.| Sequence Annotation | Location | Component / Role(s) |
| J23110 K515105 ndmA B0030 ndmC B0032 ndmD B0034 gst9 | 1,35 44,68 69,1124 1133,1147 1154,2008 2017,2029 2036,3802 3847,3858 3865,4539 | feature/promoter promoter feature/rbs ribosome_entry_site feature/cds CDS ribosome_entry_site feature/rbs CDS feature/cds feature/rbs ribosome_entry_site feature/cds CDS feature/rbs ribosome_entry_site feature/cds CDS |