Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Patrik D'haeseleer
Date created: 2015-09-17 11:00:00
Date modified: 2015-09-18 09:27:04
Ava_3856 ATP-grasp ligase
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K1839002_sequence (Version 1) |
Description
Anabaena variabilis ATCC 29413 contains a 4-gene cluster Ava_3858 to Ava_3855 that codes for a biosynthetic pathway producing the mycosporine-like amino acid (MAA) shinorine. Ava_3856 encodes and ATP-grasp like enzyme that catalyzes the addition of glycine to demethyl 4-deoxygadusol (DDG) to form mycosporine-glycine.Notes
This part uses the native cyanobacterial DNA sequence from Anabaena variabilis, with two small but important changes:1. We removed an EcoRI restriction site by by making a synonymous single-nucleotide change: GAATTC -> GAATTT.
2. We accidentally forgot to include the stop codon in our design. Luckily the BioBrick suffix does include an in-frame TAG stop codon, but if you express the gene directly from this biobrick, you can expect an extra tyrosine at the C terminal of the protein.
Source
Table S1 in [Balskus and Walsh, 2010] contains the gene locus tags and protein accession numbers for the genes in the shinorine biosynthesis gene cluster. Table S4 contains the PCR primers used for cloning the gene cluster.Ava_3857 corresponds to protein accession number ABA23461.1. The exact DNA sequence can be found in the genome sequence here:
http://www.ncbi.nlm.nih.gov/nuccore/75699950?from=4801723&to=4803099&sat=4&sat_key=105749970&report=gbwithparts
Note that the DNA sequence reported by that link is the reverse complement, since the gene is on the opposite strand in the genome sequence!
Essential references:
- Balskus EP, Walsh CT. The genetic and molecular basis for sunscreen biosynthesis in cyanobacteria. Science. 2010 Sep 24;329(5999):1653-6. http://www.ncbi.nlm.nih.gov/pmc/articles/pmid/20813918/
- Gao Q, Garcia-Pichel F. An ATP-grasp ligase involved in the last biosynthetic step of the iminomycosporine shinorine in Nostoc punctiforme ATCC 29133. J Bacteriol. 2011 Nov;193(21):5923-8. http://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21890703/
| Sequence Annotation | Location | Component / Role(s) |
| ATG start codon Ava_3856 coding region | 1,3 1,1374 | start_codon feature/start feature/cds CDS |