Team:USTC CHINA/Parts

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Parts


TD1, Transdermal peptide
Main Page-BBa_K1074000
TD1 is a short synthetic peptide(ACSSSPSKHCG)identified by in vivo phage display, facilitated efficient transdermal protein delivery through intact skin. Studies suggested that the peptide creates a transient opening in the skin barrier to enable macromolecular material to reach systemic circulation

Pgrac+RBS+SamyQ+TD1+GFP
Main Page-BBa_K1074006 This is the main circuit of our project to allow high expression of target protein(antigen,adjuvant).GFP can be substituted by various proteins via a modular PCR or standard cut/ligation method.

PHT43
PHT43 is a E.coli-B.subtilis shuttle vector allowing high-level expression of secreted proteins in B.subtilis. It is based on the strong σA-dependent promoter preceding the groE operon of B. subtilis which has been converted into an efficiently controllable (IPTGinducible)promoter by addition of the lac operator. An efficient Shine-Dalgarno (SD) sequence as well as a multiple cloning site (BamH I, Xba I, Aat II, Sma I) were also inserted. To obtain secretion of recombinant proteins, the coding region for the signal peptide of the amyQ gene encoding an α-amylase was fused to the SD sequence.

HBsAg
HBsAg is the surface antigen of the hepatitis B virus (HBV). It indicates current hepatitis B infection. Sequence and Features

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Ag85b
The antigen 85 proteins (FbpA, FbpB, FbpC) are responsible for the high affinity of mycobacteria for fibronectin, a large adhesive glycoprotein, which facilitates the attachment of M.tuberculosis to murine alveolar macrophages (AMs). They also help to maintain the integrity of the cell wall by catalyzing the transfer of mycolic acids to cell wall arabinogalactan and through the synthesis of alpha,alpha-trehalose dimycolate (TDM, cord factor). They catalyze the transfer of a mycoloyl residue from one molecule of alpha,alpha-trehalose monomycolate (TMM) to another TMM, leading to the formation of TDM. Sequence and Features

Protective Antigen Domain 4
Protective antigen (PA) is the central component of the three-part protein toxin secreted by Bacillus anthracis, the organism responsible for anthrax. Homologues of PA have been found in several spore-forming Gram-positive bacteria, and share the ability to translocate toxic enzymes into the host cytosol.The PA monomer is organised mainly into antiparallel beta-sheets and has four domains: an N-terminal domain (domain 1) containing two calcium ions and the cleavage site for activating proteases; a heptamerisation domain (domain 2) containing a large flexible loop implicated in membrane insertion; a small domain of unknown function (domain 3); and a carboxy-terminal receptor- binding domain (domain 4) [PMID: 9039918]. This entry represents domain 4 of PA, which has an immunoglobulin-like fold. Domain 4 plays a key role in cellular receptor recognition, as well as in pH- dependent pore formation[PMID: 19722284].

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NahR
Main Page- BBa_K1031610
Pre-existing Part: Main Page-BBa_J61051
BBa_K1031610 is upgraded from previous part BBa_J61051 designed by Arkin Lab. NahR is a 34 kDa transcription factor mined from Pseudomonas putida which binds to nah and sal promoters to activate transcription in response to the inducer salicylate. We constructed a Psal/NahR biosensor circuit using sfGFP as reporter gene. Adaptor of this biosensor not only improves detection efficiency but also expands detection profile.
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Parts for Biosensors

DmpR
BBa_K1031221 BBa_K1031222 BBa_K1031223 BBa_K1031224 are the RBS library of DmpR biosensor's reporter.

HbpR
BBa_K1031300 is the coding sequence of HbpR with terminators.
BBa_K1031301 and BBa_K1031302 are the RBS library of HbpR biosensor's reporter.

HcaR
BBa_K1031410 is the coding sequence of HcaR with terminators.
BBa_K1031441 BBa_K1031442 BBa_K1031443 and BBa_K1031444 are the Pc library of HbpR to tune its performance .
BBa_K1031420 and BBa_K1031421 are the RBS library of HcaR biosensor's reporter.

CapR
BBa_K1031100 is the coding sequence of CapR transcriptional factor.
BBa_K1031111 and BBa_K1031112 are basic sensor construction with different CapR expression level.
BBa_K1031113 and BBa_K1031114 are reporters with different RBS for the CapR biosensor.

HpaR
BBa_K1031510 is the coding sequence of HpaR transcriptional factor.
BBa_K1031520 and BBa_K1031521 are reporters with different RBS for the HpaR biosensor.

NahR
BBa_K1031610 is the NahR biosensor circuit.

PaaX
BBa_K1031710 is the coding sequence of PaaX with terminators.
BBa_K1031742 BBa_K1031743 and BBa_K1031744 are the Pc library of PaaX.

XylS
BBa_K1031911 is the coding sequence of XylS transcriptional factor with terminators.
BBa_K1031912 BBa_K1031913 and BBa_K1031916 are the Pc library of XylS.

XylR
BBa_K1031803 BBa_K1031804 are the RBS library of XylR biosensor's reporter.

Parts for Adaptors

NahF
BBa_K1031620 is the coding sequence of XylS transcriptional factor with terminators.
BBa_K1031621 BBa_K1031622 BBa_K1031623 BBa_K1031624 and BBa_K1031625are the Pc library of NahF.

Parts for Band-Pass Filter


BBa_K1031021 is the hybrid promoter for the reporter node, which can be activated as well as inhibited.
BBa_K1031011 is the transcriptional factor that can activate its cognate promoter, which is used as the activate node in the band-pass filter.
BBa_K1031013 BBa_K1031014 BBa_K1031015 BBa_K1031016 are RBS library to tune the expression of C1, under the control of pSal promoter, which is a crucial part for band-pass filter.

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