The localizing protein, ZRC1, a vacuolar membrane zinc transporter. C terminal fusion to ZRC1 will localize the complex to the vacuolar membrane.
The gene was codon optimized for yeast and internal restriction sites were removed for to compatibility with our Golden Gate cloning scheme.
Tadashi Nakamura, Shuhei Yasumoto, Takahiro Saka, Saya Kakuda
LacI promoter with RBS, followed by OmpA outer membrane protein and linker for Escherichia coli cell surface display.
Composite part BBa_K392001 (LacI promoter + RBS) and registry part BBa_K103006 (OmpA w/ linker).
GFP targeted to the peroxisomes of Aspergillus nidulans
Design considerations This part was designed for the Plug'n'Play with DNA assembly standard (RFC 80), therefore this part contains linkers at the 5' and 3' ends as shown on the figure below.
Expa4 plant secretion signal, localizes to extracellular space
Coding region for first 20 aminoacids of plant protein Exansin4. Secretion signal for plants (derived from A. thaliana). When fused to a protein, directs the protein to extracellular space.
Sequence was optimized for Escherichia coli and Arabidopsis thaliana codon usage.
Derived from Expansin4 from Arabidopsis thaliana(NP_181500).
EnvZ Signal Peptide and first Transmembrane Domain
For EnvZ fusion protein design. Long description will be added soon.
Source: E. coli EnvZ protein (ncbi: https://www.ncbi.nlm.nih.gov/gene/947272)
slr2016 signal sequence from cyanobacterium Synechocystis; secretes protein
The ''slr2016'' signal sequence is located at the amino terminus of slr2016 polypeptides synthesized by ''Synechocystis'' sp. PCC 6803.
Mitochondrial Leader Sequence in RFC10 standard
Mitochondrial leader sequence (MLS) helps direct protein to the mitochondria. When this sequence is cloned in frame at the N-terminus of the protein, it can target the recombinant protein to the mitochondria in mammalian cells. MLS will be removed upon translocation into the mitochondrial matrix. However, at least four additional amino acids (Ile-His-Ser-Leu) on this sequence are known to be retained on the N-terminus of the protein.
Plasmid from Invitrogen: pCMV/myc/mito (http://tools.lifetechnologies.com/content/sfs/manuals/pshooter_pcmv_man.pdf)
SV40NLS-GAL4AD-linker-PIF3 (part for GAL4/LexA based light-switchable promoter system)
CDS for a synthetic hybrid fusion protein. Consists of serveral domains. RFC10 pre and suffix
CDS for a synthetic hybrid fusion protein
SV40 nuclear localization sequence (recognized by importin alpha)
This part was designed for the Plug'n'Play with DNA assembly standard (RFC 80), therefore this part contains tails at the 5' and 3' ends as shown on the figure below. Please notice that mRFP1-NLS is not the same as BBa_K678030, BBa_K678031, or BBa_K678032.
From p71-mRFP1 of the Center for Microbial Biotechnology plasmid collection at the Technical University of Denmark. The NLS sequence was synthesized.
Mitochondrial Leader Sequence with RFC25 standard
Mitochondrial leader sequence (MLS) helps direct protein to the mitochondria. When this sequence is cloned in frame at the N-terminus of the protein, it can target the recombinant protein to the mitochondria in mammalian cells. MLS will be removed upon translocation into the mitochondrial matrix. However, at least four additional amino acids (Ile-His-Ser-Leu) on this sequence are known to be retained on the N-terminus of the protein. Since MLS always fuses with protein for function, this BioBrick is in RFC 25 standard, which facilitate protein fusion in order to introduce other protein into mitochondria.
Invitrogen: pCMV/myc/mito (http://tools.lifetechnologies.com/content/sfs/manuals/pshooter_pcmv_man.pdf)
Vector expressing eGFP from the CMV promoter with the BGH polyA tail and with a hygromycin resistance cassette
Compatible with the Plug'n'Play with DNA assembly system.
glycine-serine linker fused to B-cell receptor transmembrane region; displays protein on cell surfac
Helical single-span transmembrane region of the B-Cell-Receptor with a flexible 15 aa Linker at the N-terminus. Designed for fusion to proteins or peptides that are to be presented at the cells surface; signal peptide for membrane integration (e.g. part Bba_K157001) required at the N-terminus of the whole construct!
Between BioBrick 1.0 sites this part is flanked 5' by NgoMI(=NgoMIV) and 3' AgeI(=PinAI) to facilitate in frame cloning for protein fusions.
Gene synthesis by GeneArt, optimized for expression in homo sapiens.
myristoilation signal sequence; anchors protein to membrane
Mitochondrial targeting sequence for Aspergillus nidulans
Mitochondrial targeting sequence (MTS) for Aspergillus nidulans. A MTS is usually required for the entry of proteins and intermediary metabolites into mitochondria. The MTS is a DNA fragment encoding the N-terminal 60 amino acids of the AcuJ.
Compatible with the Plug'n'Play assembly standard. This MTS should be placed at the N-terminal of the gene of interest.
Aspergillus nidulans genomic DNA (AN6279)
N terminal signal sequence (CIIC) that targets the protein to the cellular periphery. The signal sequence isderived from the Ras protein, a signal protein located in the cell membrane that activates in response to extracellular signals for growth and differentiation
GAL4 based yeast light-switchable promoter system
BBa_K801039 (SV40NLS-GAL4AD-Linker-PIF3)
Phytochrome B (PhyB, 908 first N-terminal amino acids)
SV40 NLS (SV40 nuclear localization signal)
20 amino acids long linker (BBa_K365005)
GAL4AD (GAL4 transcriptonal activation domain)
BBa_K801040 (SV40NLS-PhyB-Linker-GAL4DBD)
Secretion tag from yeast α-factor mating pheromone (MFα1)
This part is the secretion signal peptide for the yeast α-mating factor. This signal peptide directs the secretion of the produced protein, and therefore allows for the exportation of it. This peptides are cleaved once the protein is in the lumen of the ER.
Prepro-a-factorHas a Cleavable Signal Sequence [Water et al 1987]
PKI nuclear export sequence; yeast codon optimized
yeast codon optimized PKI nuclear export sequence
J23100 Promoter with a RBS (BBa_S04423), PelB Leader sequence, Gaussia princeps luciferase, and a B0015 terminator. Allows Gaussia princeps luciferase to be transcribed and translated using a strong promoter. Luminescence can be expressed by reacting the luciferase product with coelenterazine.
This sequence is codon optimized for E.coli. Sequence contains a PelB leader sequence that allows the Gaussia princeps luciferase proteins to secreted into the periplasmic space of the cell.
Engineered from XXXXXXXX. Provided by the Bruchez lab at Carnegie Mellon.
This composite part consists of the Aga2 yeast surface display CDS linked to a flexible (Glycine3Ser)4 linker. It can be used by linking it upstream of the protein you wish to be displayed on the surface of the yeast cell. Note: Aga1 must be constitutively expressed by the yeast strain for Aga2 to anchor to the cellular surface.
This composite part was put together following RFC23, thus the scar between parts consists of the sequence ACTAGA
This part comes from the Aga2 [K416000] and Linker [K416001] parts we constructed this year.
SicP-SptP chaperone-secretion tag pair from Salmonella type III secretion system
This part is the natural implementation of the SicP chaperone (BBa_J64002) and SptP secretion signal (BBa_J64003) from the Salmonella Type III Secretion System (T3S, the Salmonella T3S coding region is referred to as Salmonella Pathogeneity Island 1 [SPI-1]). In the natural implementation the coding regions of SicP and SptP overlap out of frame. This part captures that configuration for use in creating secreted protein / chaperone pairs.
This part is not compatible with biobrick standard assembly.
SV40NLS-PhyB-linker-Gal4DBD (part for GAL4 based light switchable promoter system)
Composite part of BBa_K801030, BBa_K801030, BBa_K365005, BBa_K801035
Essential Arg for photoconvertible binding of PIF3
Essential Gly for photoconvertible binding of PIF3
Essential Cys for chromophore attachement of phycocyanobilin (PCB)
The actin targeting protein, ABP1, maintains the actin cytoskeleton. C terminally fusing a protein of interest to ABP1 will localize it to the actin.
EGFR/ErbB1 signal peptide; mediates protein transport to a translocational pore
The N-terminal signal-peptide ensures protein transport to the cytoplasm-membrane.
Part derived from the sequence of EGFR/ErbB1
When working correctly this reporter construct transcribes Aga2-eCFP from the Gal promoter. If the yeast strain is also expressing Aga1, then this fusion should be anchored onto the plasma membrane, allowing you to visualize eCFP as a halo around the yeast cell.
We considered fusing this reporter to a variety of promoters, but in the end chose pGAL because it gives a very high level of transcription and we could turn the reporter on and off
This part comes from a mixture of biobricks that were received in the 2010 initial distribution and from parts that NYU iGEM 2010 made ourselves.
Protein coding sequence for nuclear localization signal of SV40 large T antigen (amino acids: PKKKRKV).
SV40NLS-PhyB-linker-LexA (part for a LexA based light-switchable promoter system)
Composite part of BBa_K801030, BBa_K801030, BBa_K365005, BBa_K801033
LexA based yeast light-swithable promoter system
BBa_K365005 (20 amino acids long linker)
Aga2 CDS Responsible for Yeast Surface Display
The Aga2 protein is fused in frame with a protein that the user wants to display on the yeast cellular surface. Yeast cells constitutively expressing Aga1 (strain EBY100) secrete the Aga2 fusion protein, which associates with Aga1 through two disulfide bonds and quaternary structure to anchor Aga2 to the surface. The fused protein then is displayed on the surface of the yeast cell. The STOP codon at the end of the CDS was removed to allow for N- and C-terminus fusions.
We removed the STOP codon to allow for N- and C-terminal fusions, which have both been proven to occur efficiently.
This parts was biobricked from the pCTCON2 plasmid which was sent to us by Dr. D. K. Wittrup at MIT.
This secretion tag when linked to the N-terminal of the protein directs extracellular secretion of the protein. After targeting through the ER to the golgi, this "pre-pro" tag is cleaved by the KEX-2 protease resulting in secretion of protein without the aforementioned tag. To use this tag, attached it directly upstream of your proteins CDS.
There was an illegal Xbal1 site at bp 101 and we removed it through altering the primers necessary for biobricking the sequence.
This part was sent to use from Dr. Sheldon Park of the University of Buffalo.
myristoilation signal sequence fused to Tobacco Etch Virus (TEV) protease cleavage site
Signal sequence for myristoilation with proteolytic cleavage site for TEV protease.
Polypeptide containing SV40 nuclear localization sequence (SV40 NLS) for nuclear translocation
designed by oligo hybridisation based on plasmid "pGADT7 AD" from Clontech, w/ RFC25-pre and suffix for protein fusions
pSB1A3 with an C-terminal Arginine tag. This particular plasmid utilizes the BioFusion (aka Silver Lab) Standard for BioBrick assembly. Adds 10 Arginine residues to the C-terminus of a Protein (Includes Stop Codon)
pilA1 signal sequence from cyanobacterium Synechocystis; secretes protein
''pilA'' is a pilin protein known to be secreted by the cyanobacteria ''Synechocystis'' sp. PCC6803.
The ''pilA'' signal sequence is found at the amino terminal of ''pilA'' polypeptides synthesized by ''Synechocystis'' sp. PCC6803.
pSB1A3 with an N-terminal Arginine tag. This particular plasmid utilizes the BioFusion (aka Silver Lab) Standard for BioBrick assembly. Adds 10 Arginine residues to the N-terminus of a Protein.
SV40 nuclear localization sequence from SV40; yeast codon optimized
yeast codon-optimized nuclear localization sequence from simian virus 40
direct synthesis from published sequence
CD4 signal peptide; localizes to plasma membrane
Coding region for first 25 amino acids (including start codon) of CD4 receptor which is signal peptide for plasma membrane localization.
It comes from human gene for CD4 receptor.
signal sequence from Lactobacillus bulgaricus; secretes protein
This is a short signal sequence recognized as a secretion tag by several Lactobacillus strains. As the protein that is attatched to is secreted, the signal peptide is cut off. Using SignalP, the cleavage site is between residues 39 and 40 (VQA - AS). This may change depending on the following sequence. This part uses the modified Silver BioBirck prefix and suffix to allow for protein construction.
The cleavage site may be influenced by the protein that is attached to the signal peptide. To make sure that the protein does not get cleaved by mistake, four residues are added to the signal peptide sequence found by the Germond paper. Signal peptides are almost always used in conjunction with another protein so we used the modified Silver BioBrick prefix and suffix to make the construction of funcitonal fusion peptides possible
It was isolated from the 5' end of the prtB gene in Lactobacillus bulgaricus, which codes for a lactose digesting protein that is bound to the surface (Germond JE et al.; Appl Environ Microbiol. 2003). This part uses the modified Silver BioBrick prefix and suffix to allow for protein construction.
This part includes a LacY coding sequence on the backbone J61002
{rbs.pelB} The pelB leader with native rbs; assembly standard 21
This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]
OmpA outer membrane protein A fused to linker; displays proteins on cell surface
One of our basic bricks used to create fusions attached to outer membrane
PelB leader sequence; directs protein to E. coli periplasmic membrane
The pelB leader sequence is a sequence of amino acids which when attached to a protein, directs the protein to the periplasmic membrane of E. coli, where the sequence is removed by pelB peptidase. It is used to direct coat protein-antigen fusions to the cell surface.
Protein coding region for the firefly [Photinus pyralis] luciferase, codon optimized for yeast.