Promoter was amplified from genomic sequence. Rest comes from a biobrick designed by Edinburgh http://parts.igem.org/Part:BBa_I742139?title=Part:BBa_I742139
This part was created to get constitutive synthesis of yellow pigments (zeaaxnthin).
Zeaxanthin (yellow pigment) synthesis operon without PstI sites
All five genes were isolated from Pantoea ananatis (formerly Erwinia uredovora) DSMZ 30080 (ATCC 19321) More information about carotenoid synthesis may be found in these references: Diversity of Carotenoid Synthesis Gene Clusters from Environmental Enterobacteriaceae Strains, N Sedkova, APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 71(12): 8141???8146 (2005) Elucidation of Pathway the Erwinia uredovora Carotenoid Biosynthetic by Functional Analysis of Gene Products Expressed in Escherichia coli, N Misawa, et al, JOURNAL OF BACTERIOLOGY, 172(12): 6704-6712 (1990)
Zeaaxnthin, a yellow pigment, is synthesised from farnesyl disphosphate in five sequential enzymatic steps. These five enzymes are encoded by the genes CrtE, CrtB, CrtI, CrtY and CrtZ. More information on the zeaxanthin biosynthesis pathway may be found on the [http://parts.mit.edu/igem07/index.php/Edinburgh/Yoghurt/Design#Colour_Production Edinburgh Yoghurt Wiki]
Two PstI sites have been removed from CrtI gene via point mutation. All five genes were cloned so they retain their native ribosome binding sites.
Promoter expressed constitutively in a lot of strains of L. bulgaricus. Found in "Highly efficient production of the staphylococcal nuclease reporter in Lactobacillus bulgaricus governed by the promoter of the hlbA gene."
Genes were both isolated from Pantoea ananatis (formerly Erwinia uredovora) DSMZ 30080, ATCC 19321 Diversity of Carotenoid Synthesis Gene Clusters from Environmental Enterobacteriaceae Strains, N Sedkova, APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 71(12): 8141???8146 (2005) Elucidation of Pathway the Erwinia uredovora Carotenoid Biosynthetic by Functional Analysis of Gene Products Expressed in Escherichia coli, N Misawa, et al, JOURNAL OF BACTERIOLOGY, 172(12): 6704-6712 (1990)
CrtIB encodes two enzymes, which make up steps three and two in the carotenoid biosynthesis pathway respectively. CrtI encodes phytoene desaturase and CrtB encodes Phytoene synthase. Both cloned genes still have their native ribosome binding sites. More information about the carotenoid synthesis pathway may be found on the [http://parts.mit.edu/igem07/index.php/Edinburgh/Yoghurt/Design#Colour_Production Edinburgh Yoghurt Wiki]
The CrtI gene contained two PstI sites, which were both mutated out using point mutagenesis
Diversity of Carotenoid Synthesis Gene Clusters from Environmental Enterobacteriaceae Strains, N Sedkova, APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 71(12): 8141???8146 (2005) Elucidation of Pathway the Erwinia uredovora Carotenoid Biosynthetic by Functional Analysis of Gene Products Expressed in Escherichia coli, N Misawa, et al, JOURNAL OF BACTERIOLOGY, 172(12): 6704-6712 (1990) Pantoea ananatis (formerly Erwinia uredovora) DSMZ 30080 (ATCC 19321). Accession: D90087
CrtE encodes geranylgeranyl pyrophosphate synthase Enzyme converts two farnesyl diphosphate molecules into geranylgeranyl diphosphate Part of the carotenoid synthesis pathway
Ensured gene contained ribosome binding site by designing primers to include the site
from Pantoea ananatis (formerly Erwinia uredovora) DSMZ 30080 (ATCC 19321). Accession: D90087 Diversity of Carotenoid Synthesis Gene Clusters from Environmental Enterobacteriaceae Strains, N Sedkova, APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 71(12): 8141???8146 (2005) Elucidation of Pathway the Erwinia uredovora Carotenoid Biosynthetic by Functional Analysis of Gene Products Expressed in Escherichia coli, N Misawa, et al, JOURNAL OF BACTERIOLOGY, 172(12): 6704-6712 (1990)
CrtY encodes a lycopene cyclase Converts lycopene to B-carotene B-carotene is used as an orange pigment and is a vitamin A precursor
Ensured amplified gene contained native ribosome binding site
Diversity of Carotenoid Synthesis Gene Clusters from Environmental Enterobacteriaceae Strains, N Sedkova, APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 71(12): 8141???8146 (2005) Elucidation of Pathway the Erwinia uredovora Carotenoid Biosynthetic by Functional Analysis of Gene Products Expressed in Escherichia coli, N Misawa, et al, JOURNAL OF BACTERIOLOGY, 172(12): 6704-6712 (1990) from Pantoea ananatis (formerly Erwinia uredovora) DSMZ 30080 (ATCC 19321). Accession: D90087
crtZ encodes a beta-carotene hydroxylase Converts B-carotene into zeaxanthin (yellow pigment) Part of the carotenoid biosynthesis pathway
Conversion of the iGEM parts registry to SBOL2.1