Team:Lethbridge HS/Part Collection
Basic Parts:
Anthocyanin
3gt
The gene 3gt is from the anthocyanin synthesis pathway and converts the initial molecule Pelargonidin into Anthocyanin. This gene is from the organism Petunia hybrid. We have added this part to the registry as one of our new basic part submissions. Part BBa_idontknowwewillfindout
yadH
This Gene is an Escherichia coli gene that has been shown to increase the yields of anthocyanin when paired with the genes in our anthocyanin construct. It is our second original basic part submission to the registry. It is uncharacterized and is an inter-membrane transport protein. Part BBa_wowzers.
f3h
We will be useing the gene f3h as the first gene in our anthocyanin synthesis pathway, it comes from the organism Petroselinum crispum. It was added to the registry by the 2014 Darmstadt iGEM team, part BBa_K1497009. This gene will code for a protein that converts the initial molecule flavanone into dihydroflavonol.
dfr
The gene dfr is the second one in our anthocyanin synthesis pathway. We are using the biobrick part BBa_K1497010. It was added to the registry by the 2014 Darmstadt igem team.
ans
This gene is the third gene in our pathway, it converts the molecule created by dfr into pelargonidin. It is from the organism Fragaria x ananassa and was added to the registry by the 2014 Darmstadt team. It is an engineered anthocyanidin synthase. Part BBa_K1497002.
Zeaxanthin
crtY
This gene is from the organism Pantoea ananatis and is part or the carotenoid synthesis pathway. It converts the initial molecule Lycopene into the final molecule Beta-Carotene. This gene was added to the registry by Edinburgh 2007, part BBa_I742154.
crtZ
This gene is from the organism Pantoea ananatis and is our final gene in the carotenoid synthesis pathway. It was added to the registry by Edinburgh 2007 and is the part BBa_I742157.This part converts the Beat-Carotene into our final product, the pigment Zeaxanthin.
Melanin
melA
The gene melA is from the organism Rhizobium etli and was added to the registry by the Cambridge 2009 iGEM team. Part BBa_K274001.
Indigoidine
indB
This gene is from the organism Streptomyces chrmofuscus, and it is our third original basic part submission to the registry. Its is part BBa_funthings. It has been shown to increase the yeilds of Indigoidine when used with indC. It is a putative phosphatase.
indC
This gene is the gene that converts Glutamine into Indigoidine. It is from the organism Photohabdus luminescens and was added to the registry by the 2013 Heidelburg iGEM team. Part BBa_K1152013.
Additional Basic parts
T7 Promoter
The Promoter we used for our Melanin Construct is T7 promoter, This allows us to control the production of Melanin. This promoter is from the T7 bacteriophage, it is a Virus that inserts its DNA into Bacteria in order to reproduce and stay alive. This promoter works with the T7 system inside the Escherichia coli strain BL21(DE3), (Fig 1.)
(Fig 1.) This image shows the T7 Promoter system, 1.Lactose inducible promoter 2.E. coli Ribosomal Binding site. 3.The Gene for T7 RNA polymerase. 4.Terminator 5.T7 RNA polymerase. 6.T7 promoter. 7.E. coli Ribosomal Binding site. 8.The gene/genes in our construct. 9.Terminator. 10.mRNA produced by the transcription of the construct.
This shows the process that occurs with the T7 System. We induce the Lactose inducible promoter(1) with IPTG as it mimics the lactose and cannot be broken down by the cell. This allows it to continually perform transcription on the construct(1-4) embedded in the Genome of the BL21(DE3) and thusly constantly produce the T7 RNA polymerase(5) encoded by the gene(3).This T7 RNA Polymerase(5) attaches to the T7 Promoter(6) and starts transcription on the construct(6-9) within our plasmid psB1C3. This then creates the mRNA(10) which will undergo translation and produce the protein encoded by the gene(8) due to the E. coli RBS on the construct. Part BBa_I712074.
Ribosomal Binding Site
The RBS we have chosen for our constructs is an E. coli Ribosomal Binding Site, Part BBa_B0034
Terminator
The terminator we have chosen for our constructs is an E. coli terminator. Part BBa_B0015, it is a double terminator made up of one BBa_B0012 terminator and a BBa_B0010 terminator.
Composite Parts:
Anthocyanin Construct
This construct is being used to convert our initial molecule Eriodictyol into our final molecule Anthocyanin. The genes are:
T7 Promoter Part BBa_I712074.
RBS Part BBa_B0034.
f3h Part BBa_K1497009
dfr Part BBa_K1497010
ans Part BBa_K1497002
3gt Part BBa_waitingforsubmission
yadH Part BBa_alsowaiting
Terminator Part BBa_B0015.
These genes will all be used in the cell to produce Anthocyanin out of Eriodictyol. The yadH has been shown to increase yields even though it is not directly involved in the pathway. The 3gt as well as yad H are two of our original basic part submissions. This Construct is our composite part submission BBa_anthocyanincompsitepart.
Zeaxanthin Construct
This construct is made up of the following genes:
T7 Promoter Part BBa_I712074.
RBS Part BBa_B0034.
crtY Part BBa_I742154.
crtZ Part BBa_I742157.
Terminator Part BBa_B0015.
These parts all come together in our composite part BBa_Zeaxanthinconstruct. This construct is in the plasmid psB1C3, and will convert our initial molecule Lycopene into our final product, the pigment Zeaxanthin.
Indigoidine Construct
Our Indigoidine construct is made up of:
T7 Promoter PartBBa_I712074.
RBS Part BBa_B0034.
indB Part BBa_waiting for actual part submission.
indC Part BBa_K1152013
Terminator Part BBa_B0015.
These two genes come together to convert our initial molecule Glutamine into Indigoidine. The indB has been shown to increase the yields of Indigoidine as well as it is one of our original basic part submissions. This construct is part BBa_idkyet
Melanin Construct
The parts used in our Melanin construct are as follows:
T7 Promoter PartBBa_I712074.
RBS Part BBa_B0034.
MelA Part BBA_K274001.
Terminator Part BBa_B0015.
This construct allows us to make Melanin out of L-Tyrosine. it is Part BBa_waitandsee
