BBa_K1627006

BBa_K1627006 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K1627006
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
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K1627006/1