Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Alejandro Gutierrez
Date created: 2015-09-09 11:00:00
Date modified: 2015-09-10 04:37:34
Device to demethylate caffeine
| Types | DnaRegion |
| Roles | engineered_region Device |
| Sequences | BBa_K1627006_sequence (Version 1) |
Description
This part contains genes that allow for the demethylation of caffeine. The part contains a synthetic operon consisting of ndmA, ndmB, 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 caffeine in a solution. By growing the ∆guaB strain with this device in a minimal media with different known concentrations of caffeine, a growth curve can be made that correlates culture growth with concentration of caffeine. Then if the strain is grown in a minimal media with an unknown concentration of caffeine, the growth can be compared to the growth curve to calculate the initial concentration of caffeine in the media.
Notes
These genes were amplified from the pDCAF3 plasmid (BBa_K734000). A PstI restriction site was removed from the ndmA gene and an EcoRI restriction site was removed from ndmB during the cloning process. The genes were cloned into the pSB1C3 backbone using BioBrick Standard Assembly.Source
The ndmA, ndmB, 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 K082001 ndmB B0030 ndmC B0032 ndmD B0034 gst9 | 1,35 44,68 69,1124 1133,1147 1154,2221 2230,2240 2251,3105 3114,3126 3133,4899 4944,4955 4962,5636 | promoter feature/promoter feature/rbs ribosome_entry_site CDS feature/cds ribosome_entry_site feature/rbs CDS feature/cds ribosome_entry_site feature/rbs feature/cds CDS ribosome_entry_site feature/rbs CDS feature/cds feature/rbs ribosome_entry_site CDS feature/cds |