Team:Slovenia/Part Collection

Part collection

 Part Collections

 Mechanosensing Collection

Our BioBricks can work together as a coordinated system, but can also be divided into four smaller collections as mentioned above.

Our Mechanosensing Collection is composed of mechanosensitive ion channels and coding parts for protein gas vesicles, which enhance the sensitivity of mammalian cells to ultrasound or other mechanical stimuli, as well as a calcium influx measurement device based on the formation of a complex between calmodulin and M13 peptide with split luciferase, which results in the luminescence upon calcium influx.

Construct Biobrick number
MscS:HA BBa_K1965000
TRPC1:Myc BBa_K1965001
P3:FAStm:HA:TRPC1:Myc BBa_K1965002
FLAG:GvpC BBa_K1965003
Au1:GvpA BBa_K1965004
nLuc:M13 BBa_K1965014
CaM(E104Q):cLuc BBa_K1965015
nTEV:M13 BBa_K1965016
CaM(E104Q):Ctev BBa_K1965017
CaM(E31Q,E104Q):cLuc BBa_K1965018
 Touchpaint Collection

A Touchpaint Collection is a subset of the Mechanosensing Collection that enables to convert mechanical stimulus of mammalian cells into light. This set contains the parts to enhance the sensitivity of cells to mechanical stimulus and the luciferase reporter. While this collection has been used to draw on cells it has many other uses, such as detection of the shear flow, ultrasound and other types of direct and indirect mechanical stress.

Construct Biobrick number
MscS:HA BBa_K1965000
FLAG:GvpC BBa_K1965003
Au1:GvpA BBa_K1965004
nLuc:M13 BBa_K1965014
CaM(E104Q):cLuc BBa_K1965015

 Orthogonal Protease Collection

Our Orthogonal Protease Collection (nominated for Best Collection) is composed of a set of site-specific proteases that recognize different 7-aminoacid residue motif targets (PPVp, SuMMVp, SbMVp and TEVp). The proteases were tested and are deposited as single chain coding sequences and as split protein fragments for light- and chemically-inducible reconstitution. The collection also contains the corresponding cyclic luciferase reporters for each of the proteases. We determined that the new proteases are even more active than the TEVp that has been the standard in biotechnology for several decades. We demonstrated that the split versions of all four proteases can be activated by different external stimuli, such as small molecules or light, demonstrating the robustness and versatility of the system. These proteases have been used to design signaling pathways and logic circuits in mammalian cells operating at the post-translational level.

Construct Biobrick number
His:CRY:Myc:nLuc BBa_K1965007
His:CIBN:cLuc:HA BBa_K1965008
Myc:TEVP:HA BBa_K1965009
fLUC:TEVs BBa_K1965010
His:CRY:Myc:nTEV BBa_K1965019
His:CIBN:cTEV:HA BBa_K1965020
erTEVp BBa_K1965024
PPVp BBa_K1965025
FKBP:cPPVp BBa_K1965026
FRB:nPPVp BBa_K1965027
cycLuc_SbMVs BBa_K1965031
FKBP:cSbMVp:HA BBa_K1965032
Myc:FRB:nSbMVp BBa_K1965033
cycLuc_SuMMVs BBa_K1965034
FKBP:SuMMVp:HA BBa_K1965035
Myc:FRB:SuMMVp BBa_K1965036
cycLuc_TEVs BBa_K1965037
FKBP:cTEV BBa_K1965038
FRB:nTEVp BBa_K1965039
SbMVp BBa_K1965040
SuMMVp BBa_K1965041
cycLuc_PPVs BBa_K1965042
CRY:nPPV BBa_K1965043
CIB:cPPV BBa_K1965044

 Protease-based Logic Collection

Our Protease-based Logic Collection is composed of sets of parallel, antiparallel and destabilized coiled coils in fusion with parts of split luciferase. These constructs represent a tool for constructing logic circuits in mammalian cells operating at the post-translational level. The input signals for our logic operations is proteolytic activity of selected orthogonal proteases, representing constitutive logic gates, or split TEV and PPV proteases on inducible systems, representing inducible logic gates.

Construct Biobrick number
P3:cLuc BBa_K1965005
nLuc:AP4 BBa_K1965006
P5:GS10:cLuc:HA BBa_K1965011
P7:GS10:cLuc:HA BBa_K1965012
P9:GS10:cLuc:HA BBa_K1965013
nLuc:AP4:TEVs:P3mS BBa_K1965021
nLuc:AP4:TEVs:P3mS-2A BBa_K1965022
Myc:nLuc:GS10:AP6 BBa_K1965023
cLuc:A:HA BBa_K1965045
Myc:B:nLuc BBa_K1965046
cLuc:A:PPVs:B'2a:HA BBa_K1965047
Myc:A':TEVs:B:nLuc BBa_K1965048
P3:GS6:PPVs:GS6:cLuc:HA BBa_K1965049
P5:GS6:PPVs:GS6:cLuc:HA BBa_K1965050
Myc:nLuc:GS6:TEVs:GS6:AP4 BBa_K1965051

 FastER Secretion Collection

Our FastER Secretion Collection is composed of parts coding for the TagRFP reporter with different ER retention signals, attached to the reporter via a TEV cleavage site. In this way, a protein can aggregate in the ER and be secreted upon induction by proteolytic cleavage with one of our inducible split TEV proteases without having to wait for the transcription and translation of the protein.

Construct Biobrick number
ss:TagRFP:AU1:TEVs:KDEL BBa_K1965028
ss-TagRFP:AU1:furS:TM:TEVs BBa_K1965029
ss:TagRFPA:U1:furS:TM:3xTEVs:KKMP BBa_K1965030