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 08:29:33
Ava_3858 demethyl 4-deoxygadusol (DDG) synthase
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K1839000_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. Genes Ava_3858 and Ava_3857 encode demethyl 4-deoxygadusol (DDG) synthase and O-methyltransferase (O-MT), respectively, which transform sedoheptulose-7-phosphate (SHP) from the Calvin-Benson-Bassham cycle into 4-deoxygadusol (4-DG), the core structure of mycosporines.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 changing the T in position 114 to a C: GAATTC -> GAACTC.
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_3858 corresponds to protein accession number ABA23463.1. The exact DNA sequence can be found in the genome sequence here:
http://www.ncbi.nlm.nih.gov/nuccore/75699950?from=4804128&to=4805360&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!
| Sequence Annotation | Location | Component / Role(s) |
| ATG start codon Ava_3858 coding region | 1,3 1,1230 | feature/start start_codon feature/cds CDS |