//cds/enzyme/protease
type: Collection
persistent identity: cds enzyme protease
display id: cds_enzyme_protease
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
owned by: james
owned by: myers
member: BBa_K116601
member: subE
member: BBa_K772002
member: TEV
member: subE_Y77W
member: MlrAa
member: pelB+subE
member: BBa_K772003
member: BBa_M1436
member: BBa_I712020
member: MlrA
member: subE_Y77X
member: BBa_M1437
member: ExpSubE
member: BBa_K533003
member: BBa_K1378001
member: BBa_K1319004
member: BBa_K1680011
member: BBa_K1907002
member: subE_S221Y
member: BBa_I712078
member: BBa_K772001
member: BBa_K772100
member: subE_S221X
member: HIV-1
member: BBa_K1639008

MlrAa
type: ComponentDefinition
persistent identity: BBa K1907006
display id: BBa_K1907006
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Microcystinase (Mlra) + mating factor alpha tag, for S. cerevisiae
creator: Pihla Savola
created: 2016-10-18T11:00:00Z
modified: 2016-10-19T01:33:54Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2373_
m user id: 29559
owner id: 29693
owning group id: 9
status: status/Planning
bookmark: false
mutable description: --
mutable notes: --
mutable provenance: --
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: Mating alpha tag
sequence annotation: MlrA
sequence annotation: linker
sequence annotation: His-tag
sequence: https://synbiohub.org/public/igem/BBa_K1907006_sequence/1

Mating alpha tag
type: SequenceAnnotation
persistent identity: annotation2522189
display id: annotation2522189
version: 1
top level: MlrAa
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1907006/annotation2522189/range2522189/1

https://synbiohub.org/public/igem/BBa_K1907006/annotation2522189/range2522189/1
type: Range
persistent identity: range2522189
display id: range2522189
version: 1
direction: direction/forward
top level: MlrAa
start: 1
end: 267
orientation: inline

MlrA
type: SequenceAnnotation
persistent identity: annotation2522196
display id: annotation2522196
version: 1
top level: MlrAa
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1907006/annotation2522196/range2522196/1

https://synbiohub.org/public/igem/BBa_K1907006/annotation2522196/range2522196/1
type: Range
persistent identity: range2522196
display id: range2522196
version: 1
direction: direction/forward
top level: MlrAa
start: 268
end: 1275
orientation: inline

linker
type: SequenceAnnotation
persistent identity: annotation2522192
display id: annotation2522192
version: 1
top level: MlrAa
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1907006/annotation2522192/range2522192/1

https://synbiohub.org/public/igem/BBa_K1907006/annotation2522192/range2522192/1
type: Range
persistent identity: range2522192
display id: range2522192
version: 1
direction: direction/forward
top level: MlrAa
start: 1276
end: 1284
orientation: inline

His-tag
type: SequenceAnnotation
persistent identity: annotation2522210
display id: annotation2522210
version: 1
top level: MlrAa
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1907006/annotation2522210/range2522210/1

https://synbiohub.org/public/igem/BBa_K1907006/annotation2522210/range2522210/1
type: Range
persistent identity: range2522210
display id: range2522210
version: 1
direction: direction/forward
top level: MlrAa
start: 1285
end: 1302
orientation: inline

BBa_K772100
type: ComponentDefinition
persistent identity: BBa K772100
display id: BBa_K772100
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: EpCAM Detection Device
creator: Mustafa Elitok
created: 2012-09-25T11:00:00Z
modified: 2015-05-08T01:13:15Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _1024_
m user id: 0
owner id: 9573
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: true
mutable description: This gene sequence includes both of EpCAM Binding Docks; BBa_K772001 and BBa_K772002. Thus, this composite part is the main component of CTC (Circulating Tumor Cell) detection system. Docks include an EpCAM (Epithelial Cell Adhesion Molecule) antigen binding domain, OmpA transmembrane protein (dock) and TEV protease C or N terminus. Hopefully, after binding to the EpCAM antigen, TEV protease domains will fix together to form the full structure in order to activate the signalling pathway. (For more information: BBa_K772003) EpCAM is an natural antigen which is found high significantly in the cases of some breast and colorectal cancer subtypes. On the upstream region of protein sequences, there are a constitutive pLac promoter and appropriate RBS. To see how this part works: Experience.
mutable notes: There are two protein sequences between domains and the dock: One between the binding domain (antibody) and dock and one between the domain of TEV protease.
mutable provenance: CD225 (EpCAM antibody) is commonly used for the test of presence of EpCAM in the laboratories. The sequences of OmpA and TEV protease terminuses are acquired from the Registry.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Project
component: BBa_K772002
component: BBa_K772001
sequence annotation: BBa_K772002
sequence annotation: BBa_K772001
sequence: https://synbiohub.org/public/igem/BBa_K772100_sequence/1

BBa_K772002
type: Component
persistent identity: component2211348
display id: component2211348
version: 1
top level: BBa_K772100
access: public
definition: BBa_K772002

BBa_K772001
type: Component
persistent identity: component2211341
display id: component2211341
version: 1
top level: BBa_K772100
access: public
definition: BBa_K772001

BBa_K772002
type: SequenceAnnotation
persistent identity: annotation2211348
display id: annotation2211348
version: 1
top level: BBa_K772100
location: https://synbiohub.org/public/igem/BBa_K772100/annotation2211348/range2211348/1
component: BBa_K772002

https://synbiohub.org/public/igem/BBa_K772100/annotation2211348/range2211348/1
type: Range
persistent identity: range2211348
display id: range2211348
version: 1
direction: direction/reverse
top level: BBa_K772100
start: 1168
end: 2305
orientation: inline

BBa_K772001
type: SequenceAnnotation
persistent identity: annotation2211341
display id: annotation2211341
version: 1
top level: BBa_K772100
location: https://synbiohub.org/public/igem/BBa_K772100/annotation2211341/range2211341/1
component: BBa_K772001

https://synbiohub.org/public/igem/BBa_K772100/annotation2211341/range2211341/1
type: Range
persistent identity: range2211341
display id: range2211341
version: 1
direction: direction/reverse
top level: BBa_K772100
start: 1
end: 1159
orientation: inline

HIV-1
type: ComponentDefinition
persistent identity: BBa I712667
display id: BBa_I712667
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: HIV-1 aspartyl protease
creator: Rok Gaber
created: 2007-10-23T11:00:00Z
modified: 2015-08-31T04:07:47Z
discontinued: false
dominant: false
experience: experience/None
group u list: _130_
m user id: 0
owner id: 2367
owning group id: 9
sample status: In stock
status: status/Available
bookmark: false
mutable description: HIV-1 protease is an aspartyl protease that is essential for the life-cycle of HIV and cleaves proteins at specific aminoacid sequence.
mutable provenance: HIV
owned by: james
owned by: myers
star: true
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: STOP
sequence annotation: HIV protease
sequence annotation: START
sequence: https://synbiohub.org/public/igem/BBa_I712667_sequence/1

STOP
type: SequenceAnnotation
persistent identity: annotation1953713
display id: annotation1953713
version: 1
top level: HIV-1
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_I712667/annotation1953713/range1953713/1

https://synbiohub.org/public/igem/BBa_I712667/annotation1953713/range1953713/1
type: Range
persistent identity: range1953713
display id: range1953713
version: 1
direction: direction/forward
top level: HIV-1
start: 301
end: 303
orientation: inline

HIV protease
type: SequenceAnnotation
persistent identity: annotation1953711
display id: annotation1953711
version: 1
top level: HIV-1
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_I712667/annotation1953711/range1953711/1

https://synbiohub.org/public/igem/BBa_I712667/annotation1953711/range1953711/1
type: Range
persistent identity: range1953711
display id: range1953711
version: 1
direction: direction/forward
top level: HIV-1
start: 1
end: 303
orientation: inline

START
type: SequenceAnnotation
persistent identity: annotation1953712
display id: annotation1953712
version: 1
top level: HIV-1
role: SO:0000318
role: feature/start
location: https://synbiohub.org/public/igem/BBa_I712667/annotation1953712/range1953712/1

https://synbiohub.org/public/igem/BBa_I712667/annotation1953712/range1953712/1
type: Range
persistent identity: range1953712
display id: range1953712
version: 1
direction: direction/forward
top level: HIV-1
start: 1
end: 3
orientation: inline

subE
type: ComponentDefinition
persistent identity: BBa K2020000
display id: BBa_K2020000
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: subtilisin E gene
creator: Nicola Freyer, Lea Steinbeck, Svenja Meyer, Alexander Deitert
created: 2016-10-03T11:00:00Z
modified: 2016-10-04T07:43:31Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2487_
m user id: 30763
owner id: 30763
owning group id: 9
status: status/Planning
bookmark: false
mutable description: This gene can be used to express subtilisin E in Escherichia coli. Subtilisin E is an alkaline serine protease which non-specifically deigests proteins. To use this part a suitable leader sequence has to be placed in front as the sequence of this BioBrick does not contain a start codon.
mutable notes: The sequence from B. subtilis was codon optimized for E. coli with the DNA and protein sequence analysis software "Geneious" and additionally with the "Codon Optimization Tool" from IDT.
mutable provenance: The sequence wasoriginally obtained from a wildtype Bacillus subtilis and was ordered from IDT.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Other
role: SO:0000110
sequence annotation: propeptide
sequence annotation: subtilisin E
sequence: https://synbiohub.org/public/igem/BBa_K2020000_sequence/1

propeptide
type: SequenceAnnotation
persistent identity: annotation2485808
display id: annotation2485808
version: 1
top level: subE
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K2020000/annotation2485808/range2485808/1

https://synbiohub.org/public/igem/BBa_K2020000/annotation2485808/range2485808/1
type: Range
persistent identity: range2485808
display id: range2485808
version: 1
direction: direction/forward
top level: subE
start: 1
end: 230
orientation: inline

subtilisin E
type: SequenceAnnotation
persistent identity: annotation2485809
display id: annotation2485809
version: 1
top level: subE
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K2020000/annotation2485809/range2485809/1

https://synbiohub.org/public/igem/BBa_K2020000/annotation2485809/range2485809/1
type: Range
persistent identity: range2485809
display id: range2485809
version: 1
direction: direction/forward
top level: subE
start: 231
end: 1059
orientation: inline

pelB+subE
type: ComponentDefinition
persistent identity: BBa K2020001
display id: BBa_K2020001
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: subtilisin E gene with leader sequence pelB
creator: Nicola Freyer, Lea Steinbeck, Svenja Meyer, Alexander Deitert
created: 2016-10-03T11:00:00Z
modified: 2016-10-04T07:47:20Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2487_
m user id: 30763
owner id: 30763
owning group id: 9
status: status/Planning
bookmark: false
mutable description: This composite part consists of the leader sequence pelB (BBa_J32015) and the subtilisin E gene (BBa_K2020000). It can be used to express subtilisin E in Escherichia coli and simultaneously secret the enzyme into the periplasm of the cell. Subtilisin E is an alkaline serine protease which non-specifically digests proteins.
mutable notes: The sequence of the subtilisin E gene from Bacillus subtilis was codon optimized for E. coli with the DNA and protein sequence analysis software "Geneious" and additionally with the "Codon Optimization Tool" from IDT. Moreover, the leader sequence pelB begins with a start odon and was placed in front of the subtilisin E gene to generate a functional coding sequence that can be expressed in E. coli.
mutable provenance: The sequence of the signal peptide pelB was obtained through the Registry (BBa_J32015: http://parts.igem.org/Part:BBa_J32015). Together with the subtilisin E gene (BBa_K2020000: http://parts.igem.org/Part:BBa_K2020000) it was ordered from IDT.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
component: BBa_K2020000
component: BBa_J32015
sequence annotation: BBa_K2020000
sequence annotation: BBa_J32015
sequence: https://synbiohub.org/public/igem/BBa_K2020001_sequence/1

BBa_K2020000
type: Component
persistent identity: component2485824
display id: component2485824
version: 1
top level: pelB+subE
access: public
definition: subE

BBa_J32015
type: Component
persistent identity: component2485821
display id: component2485821
version: 1
top level: pelB+subE
access: public
definition: https://synbiohub.org/public/igem/BBa_J32015/1

BBa_K2020000
type: SequenceAnnotation
persistent identity: annotation2485824
display id: annotation2485824
version: 1
top level: pelB+subE
location: https://synbiohub.org/public/igem/BBa_K2020001/annotation2485824/range2485824/1
component: BBa_K2020000

https://synbiohub.org/public/igem/BBa_K2020001/annotation2485824/range2485824/1
type: Range
persistent identity: range2485824
display id: range2485824
version: 1
direction: direction/reverse
top level: pelB+subE
start: 67
end: 1125
orientation: inline

BBa_J32015
type: SequenceAnnotation
persistent identity: annotation2485821
display id: annotation2485821
version: 1
top level: pelB+subE
location: https://synbiohub.org/public/igem/BBa_K2020001/annotation2485821/range2485821/1
component: BBa_J32015

https://synbiohub.org/public/igem/BBa_K2020001/annotation2485821/range2485821/1
type: Range
persistent identity: range2485821
display id: range2485821
version: 1
direction: direction/reverse
top level: pelB+subE
start: 1
end: 66
orientation: inline

BBa_M1436
type: ComponentDefinition
persistent identity: BBa M1436
display id: BBa_M1436
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Pepsinogen from Homo sapiens optimized for E. coli
creator: Trevor Russell Ballard
created: 2013-04-24T11:00:00Z
modified: 2015-05-08T01:13:56Z
discontinued: false
dominant: false
group u list: _768_
m user id: 0
owner id: 17280
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: Pepsinogen is the precursor protein for the enzyme pepsin, one of the primary digestive enzymes located in the human stomach. Pepsinogen activates into pepsin in the presence of HCl and other pepsinogen molecules. It is a protease, being used in the human digestive system to digest proteins. This sequence has been codon optimized for expression in E. coli.
mutable notes: Codon optimized for expression in E. coli using Gene Designer from DNA 2.0.
mutable provenance: http://www.uniprot.org/uniprot/P0DJD7
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence: https://synbiohub.org/public/igem/BBa_M1436_sequence/1

BBa_M1437
type: ComponentDefinition
persistent identity: BBa M1437
display id: BBa_M1437
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Enteropeptidase from Homo sapiens optimized for E. coli
creator: Trevor Russell Ballard
created: 2013-04-24T11:00:00Z
modified: 2015-05-08T01:13:56Z
discontinued: false
dominant: false
group u list: _768_
m user id: 0
owner id: 17280
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: Enteropeptidase is an enzyme secreted in the small intestine necessary for digestion in humans. It is necessary for trypsin activation and acts as a serine protease.
mutable notes: Optimized for use in E. coli.
mutable provenance: http://www.uniprot.org/uniprot/P98073
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence: https://synbiohub.org/public/igem/BBa_M1437_sequence/1

subE_S221Y
type: ComponentDefinition
persistent identity: BBa K2020003
display id: BBa_K2020003
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: mutated expression system for subtilisin E in E. coli (S221Y)
creator: Nicola Freyer, Lea Steinbeck, Svenja Meyer, Alexander Deitert
created: 2016-10-04T11:00:00Z
modified: 2016-10-05T12:05:25Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2487_
m user id: 30763
owner id: 30763
owning group id: 9
status: status/Planning
bookmark: false
mutable description: This composite part consists of the promoter BBa_R0010, the ribosome binding site BBa_B0034, the newly created BioBrick part BBa_K2020001 and the terminator BBa_B0010. BioBrick BBa_K2020001 is a composite part itself and includes the secretion tag pelB (BBa_J32015) and a subtilisin E gene optimized for ''Escherichia coli'' codon usage (BBa_K2020000). Once introduced into ''E. coli'', this BioBrick is able to produce an inactive version of subtilisin E and simultaneously secret the enzyme into the periplasm of the cell. Subtilisin E is an alkaline serine protease which non-specifically digests proteins. By performing a site-directed mutagenesis, serine in the catalytic triade of the enzyme was exchanged against tyrosine. Therefore, the enzyme looses its proteolytic activity.
mutable notes: The sequence of the subtilisin E gene from ''Bacillus subtilis'' was codon optimized for ''E. coli'' with the DNA and protein sequence analysis software "Geneious" and additionally with the "Codon Optimization Tool" from IDT. The sequence was partly ordered from IDT (BBa_K2020001 + BBa_B0010) and then cloned into BBa_J04500, a protein expression backbone which already carries the LacI promoter BBa_R0010 and the ribosome binding site BBa_B0034. Afterwards, a mutation in the active site of the enzyme was introduced by performing a site-directed mutagenesis. The codon AGC of serine221 was substituted with TAC which codes for tyrosine.
mutable provenance: The sequences of the BioBrick parts were obtained through the Registry. {| class="wikitable" style="text-align:center" |- ! part !! colspan="2"| BioBrick No. |- | LacI promoter || colspan="2"| BBa_R0010 |- | RBS || colspan="2"| BBa_B0034 |- | rowspan="2" | CDS || colspan="2"| BBa_K2020001 |- | BBa_J32015 || BBa_K2020000 |- | terminator || colspan="2"| BBa_B0010 |- |} The construct was ordered at IDT.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Composite
sequence annotation: AGC1182TAC
sequence annotation: BBa_B0010
sequence annotation: subtilisin E
sequence annotation: BBa_B0034
sequence annotation: LacI
sequence annotation: PelB
sequence annotation: propeptide
sequence: https://synbiohub.org/public/igem/BBa_K2020003_sequence/1

AGC1182TAC
type: SequenceAnnotation
persistent identity: annotation2486161
display id: annotation2486161
version: 1
top level: subE_S221Y
role: SO:0001059
role: feature/mutation
location: https://synbiohub.org/public/igem/BBa_K2020003/annotation2486161/range2486161/1

https://synbiohub.org/public/igem/BBa_K2020003/annotation2486161/range2486161/1
type: Range
persistent identity: range2486161
display id: range2486161
version: 1
direction: direction/forward
top level: subE_S221Y
start: 1182
end: 1184
orientation: inline

BBa_B0010
type: SequenceAnnotation
persistent identity: annotation2486160
display id: annotation2486160
version: 1
top level: subE_S221Y
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K2020003/annotation2486160/range2486160/1

https://synbiohub.org/public/igem/BBa_K2020003/annotation2486160/range2486160/1
type: Range
persistent identity: range2486160
display id: range2486160
version: 1
direction: direction/forward
top level: subE_S221Y
start: 1360
end: 1439
orientation: inline

subtilisin E
type: SequenceAnnotation
persistent identity: annotation2486159
display id: annotation2486159
version: 1
top level: subE_S221Y
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K2020003/annotation2486159/range2486159/1

https://synbiohub.org/public/igem/BBa_K2020003/annotation2486159/range2486159/1
type: Range
persistent identity: range2486159
display id: range2486159
version: 1
direction: direction/forward
top level: subE_S221Y
start: 523
end: 1351
orientation: inline

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2486156
display id: annotation2486156
version: 1
top level: subE_S221Y
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K2020003/annotation2486156/range2486156/1

https://synbiohub.org/public/igem/BBa_K2020003/annotation2486156/range2486156/1
type: Range
persistent identity: range2486156
display id: range2486156
version: 1
direction: direction/forward
top level: subE_S221Y
start: 209
end: 220
orientation: inline

LacI
type: SequenceAnnotation
persistent identity: annotation2486155
display id: annotation2486155
version: 1
top level: subE_S221Y
role: SO:0000167
role: feature/promoter
location: https://synbiohub.org/public/igem/BBa_K2020003/annotation2486155/range2486155/1

https://synbiohub.org/public/igem/BBa_K2020003/annotation2486155/range2486155/1
type: Range
persistent identity: range2486155
display id: range2486155
version: 1
direction: direction/forward
top level: subE_S221Y
start: 1
end: 200
orientation: inline

PelB
type: SequenceAnnotation
persistent identity: annotation2486157
display id: annotation2486157
version: 1
top level: subE_S221Y
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K2020003/annotation2486157/range2486157/1

https://synbiohub.org/public/igem/BBa_K2020003/annotation2486157/range2486157/1
type: Range
persistent identity: range2486157
display id: range2486157
version: 1
direction: direction/forward
top level: subE_S221Y
start: 227
end: 292
orientation: inline

propeptide
type: SequenceAnnotation
persistent identity: annotation2486158
display id: annotation2486158
version: 1
top level: subE_S221Y
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K2020003/annotation2486158/range2486158/1

https://synbiohub.org/public/igem/BBa_K2020003/annotation2486158/range2486158/1
type: Range
persistent identity: range2486158
display id: range2486158
version: 1
direction: direction/forward
top level: subE_S221Y
start: 293
end: 522
orientation: inline

BBa_K533003
type: ComponentDefinition
persistent identity: BBa K533003
display id: BBa_K533003
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: OmpA-HIV aspartyl protease
creator: ZHANG Yunxiao
created: 2011-09-28T11:00:00Z
modified: 2015-05-08T01:12:36Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _699_
m user id: 0
owner id: 6884
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: true
mutable description: OmpA is used to present HIV aspartyl protease outside the E. Coli membrane so that it can cut other proteins with the specific sequence outside in the media.
mutable notes: Since HIV protease is a dimer, dimerization is a great concern to its function. Function of this protease is tested in BL21.
mutable provenance: HIV protease sequence is from former part BBa_I712667. OmpA sequence is from BL21(DE3) e. Coli strain.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Device
role: SO:0000804
sequence annotation: OmpA
sequence annotation: T7
sequence annotation: START
sequence annotation: STOP
sequence annotation: HIV protease
sequence annotation: rbs
sequence annotation: BBa_I712667
sequence annotation: lacI
sequence: https://synbiohub.org/public/igem/BBa_K533003_sequence/1

OmpA
type: SequenceAnnotation
persistent identity: annotation2150441
display id: annotation2150441
version: 1
top level: BBa_K533003
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K533003/annotation2150441/range2150441/1

https://synbiohub.org/public/igem/BBa_K533003/annotation2150441/range2150441/1
type: Range
persistent identity: range2150441
display id: range2150441
version: 1
direction: direction/forward
top level: BBa_K533003
start: 87
end: 632
orientation: inline

T7
type: SequenceAnnotation
persistent identity: annotation2150439
display id: annotation2150439
version: 1
top level: BBa_K533003
role: SO:0000167
role: feature/promoter
location: https://synbiohub.org/public/igem/BBa_K533003/annotation2150439/range2150439/1

https://synbiohub.org/public/igem/BBa_K533003/annotation2150439/range2150439/1
type: Range
persistent identity: range2150439
display id: range2150439
version: 1
direction: direction/forward
top level: BBa_K533003
start: 1
end: 18
orientation: inline

START
type: SequenceAnnotation
persistent identity: annotation2146916
display id: annotation2146916
version: 1
top level: BBa_K533003
role: SO:0000318
role: feature/start
location: https://synbiohub.org/public/igem/BBa_K533003/annotation2146916/range2146916/1

https://synbiohub.org/public/igem/BBa_K533003/annotation2146916/range2146916/1
type: Range
persistent identity: range2146916
display id: range2146916
version: 1
direction: direction/forward
top level: BBa_K533003
start: 87
end: 89
orientation: inline

STOP
type: SequenceAnnotation
persistent identity: annotation2146917
display id: annotation2146917
version: 1
top level: BBa_K533003
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K533003/annotation2146917/range2146917/1

https://synbiohub.org/public/igem/BBa_K533003/annotation2146917/range2146917/1
type: Range
persistent identity: range2146917
display id: range2146917
version: 1
direction: direction/forward
top level: BBa_K533003
start: 933
end: 935
orientation: inline

HIV protease
type: SequenceAnnotation
persistent identity: annotation2146915
display id: annotation2146915
version: 1
top level: BBa_K533003
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K533003/annotation2146915/range2146915/1

https://synbiohub.org/public/igem/BBa_K533003/annotation2146915/range2146915/1
type: Range
persistent identity: range2146915
display id: range2146915
version: 1
direction: direction/forward
top level: BBa_K533003
start: 633
end: 935
orientation: inline

rbs
type: SequenceAnnotation
persistent identity: annotation2150442
display id: annotation2150442
version: 1
top level: BBa_K533003
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K533003/annotation2150442/range2150442/1

https://synbiohub.org/public/igem/BBa_K533003/annotation2150442/range2150442/1
type: Range
persistent identity: range2150442
display id: range2150442
version: 1
direction: direction/forward
top level: BBa_K533003
start: 75
end: 79
orientation: inline

BBa_I712667
type: SequenceAnnotation
persistent identity: annotation2146918
display id: annotation2146918
version: 1
top level: BBa_K533003
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K533003/annotation2146918/range2146918/1

https://synbiohub.org/public/igem/BBa_K533003/annotation2146918/range2146918/1
type: Range
persistent identity: range2146918
display id: range2146918
version: 1
direction: direction/reverse
top level: BBa_K533003
start: 633
end: 935
orientation: inline

lacI
type: SequenceAnnotation
persistent identity: annotation2150440
display id: annotation2150440
version: 1
top level: BBa_K533003
role: SO:0000057
role: feature/operator
location: https://synbiohub.org/public/igem/BBa_K533003/annotation2150440/range2150440/1

https://synbiohub.org/public/igem/BBa_K533003/annotation2150440/range2150440/1
type: Range
persistent identity: range2150440
display id: range2150440
version: 1
direction: direction/forward
top level: BBa_K533003
start: 20
end: 44
orientation: inline

BBa_I712078
type: ComponentDefinition
persistent identity: BBa I712078
display id: BBa_I712078
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: TEV protease (C-terminal half)
creator: Rok Gaber
created: 2007-10-21T11:00:00Z
modified: 2015-08-31T04:07:46Z
discontinued: false
dominant: false
group u list: _130_
m user id: 0
owner id: 1856
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable description: C-terminal half of TEV (Tobacco Etch Virus) protease. TEV protease is a highly site-specific protease that is found in the Tobacco Etch Virus (TEV). One of the main uses of this protease is for removing affinity tags from purified proteins. But scientists developed assay for detecting transmembrane protein interactions on its base. TEV protease is splitted into halves (N-terminal and C-terminal). These halves are inactive but assembles and regain activity if they are fused with interacting transmembrane proteins. For detecting TEV protease activity another fusion protein is constructed which is composed of transmembrane region, specific TEV protease cleavage site and kind of transcription factor. When both halves of TEV protease assembles and therefore split protease regains activity it cleaves third protein and transcription factor is released from membrane. Transcription factor is later translocated to the nucleus where induces transcription of promoter with reporter gene. This is C-terminal half of this protease.
mutable notes: Synthetic gene
mutable provenance: Tobacco Etch Virus
owned by: james
owned by: myers
star: true
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: Coding
sequence annotation: STOP
sequence: https://synbiohub.org/public/igem/BBa_I712078_sequence/1

Coding
type: SequenceAnnotation
persistent identity: annotation1952299
display id: annotation1952299
version: 1
top level: BBa_I712078
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_I712078/annotation1952299/range1952299/1

https://synbiohub.org/public/igem/BBa_I712078/annotation1952299/range1952299/1
type: Range
persistent identity: range1952299
display id: range1952299
version: 1
direction: direction/forward
top level: BBa_I712078
start: 1
end: 372
orientation: inline

STOP
type: SequenceAnnotation
persistent identity: annotation1952300
display id: annotation1952300
version: 1
top level: BBa_I712078
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_I712078/annotation1952300/range1952300/1

https://synbiohub.org/public/igem/BBa_I712078/annotation1952300/range1952300/1
type: Range
persistent identity: range1952300
display id: range1952300
version: 1
direction: direction/forward
top level: BBa_I712078
start: 373
end: 375
orientation: inline

BBa_K116601
type: ComponentDefinition
persistent identity: BBa K116601
display id: BBa_K116601
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: HtlB (ftsH) coding region from E. coli
creator: Jesse Wu
created: 2008-10-29T12:00:00Z
modified: 2015-05-08T01:09:33Z
discontinued: false
dominant: false
group u list: _210_
m user id: 0
owner id: 2749
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable description: The HtlB (ftsH) coding region from E. coli. It can be used to degrade many different proteins.
mutable notes: PCR primer design
mutable provenance: E. coli K. 12 MG1655
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence: https://synbiohub.org/public/igem/BBa_K116601_sequence/1

BBa_K1680011
type: ComponentDefinition
persistent identity: BBa K1680011
display id: BBa_K1680011
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: HCV protease with linkers
creator: Katharina Sporbeck
created: 2015-09-16T11:00:00Z
modified: 2015-10-27T01:38:42Z
discontinued: false
dominant: false
group u list: _2098_
m user id: 15693
owner id: 26460
owning group id: 9
status: status/Planning
bookmark: false
mutable description: Protein coding region for the Hepatitis C virus (HCV) NS3-4A protease with N- & C-terminal linkers. Intended for combination with 145N Dronpa (Part BBa_K1680006) as described by Zhou et al. 2012. Codon optimized for Yeast.
mutable notes: Shipped in RFC25
mutable provenance: gBlock designed by Team Tuebingen
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence: https://synbiohub.org/public/igem/BBa_K1680011_sequence/1

BBa_K1907002
type: ComponentDefinition
persistent identity: BBa K1907002
display id: BBa_K1907002
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Microcystinase (MlrA) for E. coli
creator: Pihla Savola
created: 2016-10-13T11:00:00Z
modified: 2016-10-19T05:38:51Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2373_
m user id: 29560
owner id: 29693
owning group id: 9
status: status/Planning
bookmark: false
mutable description: 123
mutable notes: 123
mutable provenance: 123
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: MlrA CDS
sequence annotation: His-tag
sequence annotation: Linker
sequence: https://synbiohub.org/public/igem/BBa_K1907002_sequence/1

MlrA CDS
type: SequenceAnnotation
persistent identity: annotation2522034
display id: annotation2522034
version: 1
top level: BBa_K1907002
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1907002/annotation2522034/range2522034/1

https://synbiohub.org/public/igem/BBa_K1907002/annotation2522034/range2522034/1
type: Range
persistent identity: range2522034
display id: range2522034
version: 1
direction: direction/forward
top level: BBa_K1907002
start: 1
end: 1008
orientation: inline

His-tag
type: SequenceAnnotation
persistent identity: annotation2522044
display id: annotation2522044
version: 1
top level: BBa_K1907002
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1907002/annotation2522044/range2522044/1

https://synbiohub.org/public/igem/BBa_K1907002/annotation2522044/range2522044/1
type: Range
persistent identity: range2522044
display id: range2522044
version: 1
direction: direction/forward
top level: BBa_K1907002
start: 1017
end: 1035
orientation: inline

Linker
type: SequenceAnnotation
persistent identity: annotation2522046
display id: annotation2522046
version: 1
top level: BBa_K1907002
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1907002/annotation2522046/range2522046/1

https://synbiohub.org/public/igem/BBa_K1907002/annotation2522046/range2522046/1
type: Range
persistent identity: range2522046
display id: range2522046
version: 1
direction: direction/forward
top level: BBa_K1907002
start: 1009
end: 1017
orientation: inline

TEV
type: ComponentDefinition
persistent identity: BBa I712077
display id: BBa_I712077
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: TEV protease (N-terminal half) - entered sequence may be wrong
creator: Rok Gaber
created: 2007-10-21T11:00:00Z
modified: 2015-08-31T04:07:46Z
discontinued: false
dominant: false
experience: experience/None
group u list: _130_
m user id: 0
owner id: 1856
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable description: N-terminal half of TEV (Tobacco Etch Virus) protease. TEV protease is a highly site-specific protease that is found in the Tobacco Etch Virus (TEV). One of the main uses of this protease is for removing affinity tags from purified proteins. But scientists developed assay for detecting transmembrane protein interactions on its base. TEV protease is splitted into halves (N-terminal and C-terminal). These halves are inactive but assembles and regain activity if they are fused with interacting transmembrane proteins. For detecting TEV protease activity another fusion protein is constructed which is composed of transmembrane region, specific TEV protease cleavage site and kind of transcription factor. When both halves of TEV protease assembles and therefore split protease regains activity it cleaves third protein and transcription factor is released from membrane. Transcription factor is later translocated to the nucleus where induces transcription of promoter with reporter gene. This is N-terminal half of this protease.
mutable notes: Synthetic gene
mutable provenance: Tobacco Etch Virus
owned by: james
owned by: myers
star: true
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: Coding
sequence annotation: STOP
sequence: https://synbiohub.org/public/igem/BBa_I712077_sequence/1

Coding
type: SequenceAnnotation
persistent identity: annotation1952295
display id: annotation1952295
version: 1
top level: TEV
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_I712077/annotation1952295/range1952295/1

https://synbiohub.org/public/igem/BBa_I712077/annotation1952295/range1952295/1
type: Range
persistent identity: range1952295
display id: range1952295
version: 1
direction: direction/forward
top level: TEV
start: 1
end: 354
orientation: inline

STOP
type: SequenceAnnotation
persistent identity: annotation1952296
display id: annotation1952296
version: 1
top level: TEV
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_I712077/annotation1952296/range1952296/1

https://synbiohub.org/public/igem/BBa_I712077/annotation1952296/range1952296/1
type: Range
persistent identity: range1952296
display id: range1952296
version: 1
direction: direction/forward
top level: TEV
start: 355
end: 357
orientation: inline

ExpSubE
type: ComponentDefinition
persistent identity: BBa K2020002
display id: BBa_K2020002
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: expression system for subtilisin E in E. coli
creator: Nicola Freyer, Lea Steinbeck, Svenja Meyer, Alexander Deitert
created: 2016-10-04T11:00:00Z
modified: 2016-10-05T10:39:31Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2487_
m user id: 30763
owner id: 30763
owning group id: 9
status: status/Planning
bookmark: false
mutable description: This composite part consists of the promoter [http://parts.igem.org/Part:BBa_R0010 BBa_R0010], the RBS [http://parts.igem.org/Part:BBa_B0034 BBa_B0034], the newly created BioBrick part [http://parts.igem.org/Part:BBa_K2020001 BBa_K2020001] and the terminator [http://parts.igem.org/Part:BBa_B0010 BBa_B0010]. BioBrick BBa_K2020001 is a composite part itself and includes the secretion tag pelB ([http://parts.igem.org/Part:BBa_J32015 BBaJ32015]) and a subtilisin E gene optimized for E. coli codon usage ([http://parts.igem.org/Part:BBa_K2020000 BBa_K2020000]). Once introduced into Escherichia coli, this BioBrick is able to produce subtilisin E and simultaneously secret the enzyme into the periplasm of the cell. Subtilisin E is an alkaline serine protease which non-specifically digests proteins.
mutable notes: The sequence of the subtilisin E gene from Bacillus subtilis was codon optimized for E. coli with the DNA and protein sequence analysis software "Geneious" and additionally with the "Codon Optimization Tool" from IDT.
mutable provenance: The sequences of the BioBrick parts were obtained through the Registry. The construct was ordered at IDT. add links
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Composite
component: BBa_R0010
component: BBa_K2020001
component: BBa_B0034
component: BBa_B0010
sequence annotation: BBa_R0010
sequence annotation: BBa_K2020001
sequence annotation: BBa_B0010
sequence annotation: BBa_B0034
sequence: https://synbiohub.org/public/igem/BBa_K2020002_sequence/1

BBa_R0010
type: Component
persistent identity: component2486138
display id: component2486138
version: 1
top level: ExpSubE
access: public
definition: https://synbiohub.org/public/igem/BBa_R0010/1

BBa_K2020001
type: Component
persistent identity: component2486152
display id: component2486152
version: 1
top level: ExpSubE
access: public
definition: pelB+subE

BBa_B0034
type: Component
persistent identity: component2486146
display id: component2486146
version: 1
top level: ExpSubE
access: public
definition: https://synbiohub.org/public/igem/BBa_B0034/1

BBa_B0010
type: Component
persistent identity: component2486153
display id: component2486153
version: 1
top level: ExpSubE
access: public
definition: https://synbiohub.org/public/igem/BBa_B0010/1

BBa_R0010
type: SequenceAnnotation
persistent identity: annotation2486138
display id: annotation2486138
version: 1
top level: ExpSubE
location: https://synbiohub.org/public/igem/BBa_K2020002/annotation2486138/range2486138/1
component: BBa_R0010

https://synbiohub.org/public/igem/BBa_K2020002/annotation2486138/range2486138/1
type: Range
persistent identity: range2486138
display id: range2486138
version: 1
direction: direction/forward
top level: ExpSubE
start: 1
end: 200
orientation: inline

BBa_K2020001
type: SequenceAnnotation
persistent identity: annotation2486152
display id: annotation2486152
version: 1
top level: ExpSubE
location: https://synbiohub.org/public/igem/BBa_K2020002/annotation2486152/range2486152/1
component: BBa_K2020001

https://synbiohub.org/public/igem/BBa_K2020002/annotation2486152/range2486152/1
type: Range
persistent identity: range2486152
display id: range2486152
version: 1
direction: direction/reverse
top level: ExpSubE
start: 227
end: 1351
orientation: inline

BBa_B0010
type: SequenceAnnotation
persistent identity: annotation2486153
display id: annotation2486153
version: 1
top level: ExpSubE
location: https://synbiohub.org/public/igem/BBa_K2020002/annotation2486153/range2486153/1
component: BBa_B0010

https://synbiohub.org/public/igem/BBa_K2020002/annotation2486153/range2486153/1
type: Range
persistent identity: range2486153
display id: range2486153
version: 1
direction: direction/forward
top level: ExpSubE
start: 1360
end: 1439
orientation: inline

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2486146
display id: annotation2486146
version: 1
top level: ExpSubE
location: https://synbiohub.org/public/igem/BBa_K2020002/annotation2486146/range2486146/1
component: BBa_B0034

https://synbiohub.org/public/igem/BBa_K2020002/annotation2486146/range2486146/1
type: Range
persistent identity: range2486146
display id: range2486146
version: 1
direction: direction/reverse
top level: ExpSubE
start: 209
end: 220
orientation: inline

MlrA
type: ComponentDefinition
persistent identity: BBa K1907001
display id: BBa_K1907001
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Microcystinase (MlrA) for S. cerevisiae
creator: Pihla Savola
created: 2016-10-11T11:00:00Z
modified: 2016-10-19T12:43:58Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2373_
m user id: 29559
owner id: 29693
owning group id: 9
status: status/Planning
bookmark: false
mutable description: - - - - - - - - -
mutable notes: -- -- --
mutable provenance: -- -- --
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: Linker
sequence annotation: MlrA CDS
sequence annotation: His-tag
sequence: https://synbiohub.org/public/igem/BBa_K1907001_sequence/1

Linker
type: SequenceAnnotation
persistent identity: annotation2522063
display id: annotation2522063
version: 1
top level: MlrA
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1907001/annotation2522063/range2522063/1

https://synbiohub.org/public/igem/BBa_K1907001/annotation2522063/range2522063/1
type: Range
persistent identity: range2522063
display id: range2522063
version: 1
direction: direction/forward
top level: MlrA
start: 1009
end: 1017
orientation: inline

MlrA CDS
type: SequenceAnnotation
persistent identity: annotation2522062
display id: annotation2522062
version: 1
top level: MlrA
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1907001/annotation2522062/range2522062/1

https://synbiohub.org/public/igem/BBa_K1907001/annotation2522062/range2522062/1
type: Range
persistent identity: range2522062
display id: range2522062
version: 1
direction: direction/forward
top level: MlrA
start: 1
end: 1008
orientation: inline

His-tag
type: SequenceAnnotation
persistent identity: annotation2522064
display id: annotation2522064
version: 1
top level: MlrA
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1907001/annotation2522064/range2522064/1

https://synbiohub.org/public/igem/BBa_K1907001/annotation2522064/range2522064/1
type: Range
persistent identity: range2522064
display id: range2522064
version: 1
direction: direction/forward
top level: MlrA
start: 1018
end: 1035
orientation: inline

BBa_K772002
type: ComponentDefinition
persistent identity: BBa K772002
display id: BBa_K772002
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: EpCAM Binding Dock: N terminus
creator: Mustafa Elitok
created: 2012-09-23T11:00:00Z
modified: 2015-05-08T01:13:15Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _1024_
m user id: 0
owner id: 9573
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: true
mutable description: This gene sequence includes an EpCAM (Epithelial p? Cell Adhesion Molecule) antigen binding domain, OmpA transmembrane protein (dock) and TEV protease N terminus. It is planned to use this subpart with BBa_772, which contains only TEV protease C terminus differently instead of N terminus, in order to induce a signal reaction in the case of EpCAM presence. EpCAM is an natural antigen which is found high significantly in the cases of some breast and colorectal cancer subtypes. On the upstream region of protein sequence, there are a constitutive pLac promoter and appropiate RBS. To see how this part works: Experience.
mutable notes: There are two protein sequences between domains and the dock: One between the binding domain (antibody) and dock and one between the part of TEV protease.
mutable provenance: CD225 (EpCAM antibody) is commonly used for the test of presence of EpCAM in the laboratories. The sequences of OmpA and TEV protease terminuses are acquired from the Registry.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Translational Unit
role: SO:0000804
sequence annotation: RBS
sequence annotation: TEV Protease N terminus
sequence annotation: OmpA (Dock)
sequence annotation: CD215 antibody
sequence annotation: pCons Promoter
sequence annotation: Binding Dock
sequence: https://synbiohub.org/public/igem/BBa_K772002_sequence/1

RBS
type: SequenceAnnotation
persistent identity: annotation2203683
display id: annotation2203683
version: 1
top level: BBa_K772002
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K772002/annotation2203683/range2203683/1

https://synbiohub.org/public/igem/BBa_K772002/annotation2203683/range2203683/1
type: Range
persistent identity: range2203683
display id: range2203683
version: 1
direction: direction/forward
top level: BBa_K772002
start: 73
end: 93
orientation: inline

TEV Protease N terminus
type: SequenceAnnotation
persistent identity: annotation2203685
display id: annotation2203685
version: 1
top level: BBa_K772002
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K772002/annotation2203685/range2203685/1

https://synbiohub.org/public/igem/BBa_K772002/annotation2203685/range2203685/1
type: Range
persistent identity: range2203685
display id: range2203685
version: 1
direction: direction/forward
top level: BBa_K772002
start: 94
end: 398
orientation: inline

OmpA (Dock)
type: SequenceAnnotation
persistent identity: annotation2203682
display id: annotation2203682
version: 1
top level: BBa_K772002
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K772002/annotation2203682/range2203682/1

https://synbiohub.org/public/igem/BBa_K772002/annotation2203682/range2203682/1
type: Range
persistent identity: range2203682
display id: range2203682
version: 1
direction: direction/forward
top level: BBa_K772002
start: 399
end: 836
orientation: inline

CD215 antibody
type: SequenceAnnotation
persistent identity: annotation2203686
display id: annotation2203686
version: 1
top level: BBa_K772002
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K772002/annotation2203686/range2203686/1

https://synbiohub.org/public/igem/BBa_K772002/annotation2203686/range2203686/1
type: Range
persistent identity: range2203686
display id: range2203686
version: 1
direction: direction/forward
top level: BBa_K772002
start: 837
end: 1138
orientation: inline

pCons Promoter
type: SequenceAnnotation
persistent identity: annotation2203681
display id: annotation2203681
version: 1
top level: BBa_K772002
role: SO:0000167
role: feature/promoter
location: https://synbiohub.org/public/igem/BBa_K772002/annotation2203681/range2203681/1

https://synbiohub.org/public/igem/BBa_K772002/annotation2203681/range2203681/1
type: Range
persistent identity: range2203681
display id: range2203681
version: 1
direction: direction/forward
top level: BBa_K772002
start: 1
end: 72
orientation: inline

Binding Dock
type: SequenceAnnotation
persistent identity: annotation2203684
display id: annotation2203684
version: 1
top level: BBa_K772002
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K772002/annotation2203684/range2203684/1

https://synbiohub.org/public/igem/BBa_K772002/annotation2203684/range2203684/1
type: Range
persistent identity: range2203684
display id: range2203684
version: 1
direction: direction/forward
top level: BBa_K772002
start: 94
end: 1138
orientation: inline

BBa_K772003
type: ComponentDefinition
persistent identity: BBa K772003
display id: BBa_K772003
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: TEV protease Full Structure
creator: Mustafa Elitok
created: 2012-09-23T11:00:00Z
modified: 2015-05-08T01:13:15Z
discontinued: false
dominant: false
experience: experience/None
group u list: _1024_
m user id: 0
owner id: 9573
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: This part is synthesized for the purpose of using it whether our TEV protease based tumor detection system works correctly. TEV protease full structure pretends that its N and C terminus parts are fixed together effectively and that it is ready to cleave target sequences. In our project, there are anchor-inducer protein complexes which are planned to be cleaved by TEV protease. (The cleavage sites are used to fix these two proteins of complex.) After this reaction, it is hopefully possible to see the reporter signal which is released free after the cleavege. This signal will indicate that N and C terminuses of TEV protease are assembled by the effect of detection in the system they used. (In our case, it is the physical antibody-antigen binding effect which TEV protease parts are clustered the intracellular regiın of binding domains)
mutable notes: This part will be assembled with pLac promoter, RBS and LacI sequences in order to be regulated for the control in the assays by the addition of IPTG in the media.
mutable provenance: ?
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Translational Unit
role: SO:0000804
sequence annotation: TEV protease
sequence: https://synbiohub.org/public/igem/BBa_K772003_sequence/1

TEV protease
type: SequenceAnnotation
persistent identity: annotation2195667
display id: annotation2195667
version: 1
top level: BBa_K772003
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K772003/annotation2195667/range2195667/1

https://synbiohub.org/public/igem/BBa_K772003/annotation2195667/range2195667/1
type: Range
persistent identity: range2195667
display id: range2195667
version: 1
direction: direction/forward
top level: BBa_K772003
start: 1
end: 1044
orientation: inline

subE_S221X
type: ComponentDefinition
persistent identity: BBa K2020004
display id: BBa_K2020004
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: mutated expression system for subtilisin E in E. coli (S221X)
creator: Nicola Freyer, Lea Steinbeck, Svenja Meyer, Alexander Deitert
created: 2016-10-07T11:00:00Z
modified: 2016-10-08T11:53:10Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2487_
m user id: 30763
owner id: 30763
owning group id: 9
status: status/Planning
bookmark: false
mutable description: This composite part consists of the promoter BBa_R0010, the ribosome binding site BBa_B0034, the newly created BioBrick part BBa_K2020001 and the terminator BBa_B0010. BioBrick BBa_K2020001 is a composite part itself and includes the secretion tag pelB (BBa_J32015) and a subtilisin E gene optimized for ''Escherichia coli'' codon usage (BBa_K2020000). Once introduced into ''E. coli'', this BioBrick is able to produce an inactive version of subtilisin E and simultaneously secret the enzyme into the periplasm of the cell. Subtilisin E is an alkaline serine protease which non-specifically digests proteins. By performing a site-directed mutagenesis, serine in the catalytic triade of the enzyme was exchanged against the amber stop codon UAG. Therefore, the expression of the enzyme is interrupted. This composite part was created to integrate a non-canonical amino acid into the sequence of subtilisin E by adding an orthogonal tRNA/aminoacyl-synthetase pair that is capable of incorporating the non-canonical amino acid of choice at the UAG codon.
mutable notes: The sequence of the subtilisin E gene from ''Bacillus subtilis'' was codon optimized for ''E. coli'' with the DNA and protein sequence analysis software "Geneious" and additionally with the "Codon Optimization Tool" from IDT. The sequence was partly ordered from IDT (BBa_K2020001 + BBa_B0010) and then cloned into BBa_J04500, a protein expression backbone which already carries the LacI promoter BBa_R0010 and the ribosome binding site BBa_B0034. Afterwards, a mutation in the active site of the enzyme was introduced by performing a site-directed mutagenesis. The codon AGC of serine221 was substituted with the stop codon TAG.
mutable provenance: The sequences of the BioBrick parts were obtained through the Registry. {| class="wikitable" style="text-align:center" |- ! part !! colspan="2"| BioBrick No. |- | LacI promoter || colspan="2"| BBa_R0010 |- | RBS || colspan="2"| BBa_B0034 |- | rowspan="2" | CDS || colspan="2"| BBa_K2020001 |- | BBa_J32015 || BBa_K2020000 |- | terminator || colspan="2"| BBa_B0010 |- |} The construct was ordered at IDT.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Composite
sequence annotation: AGC1182TAG
sequence annotation: propeptide
sequence annotation: subtilisin E
sequence annotation: BBa_B0010
sequence annotation: BBa_B0034
sequence annotation: LacI
sequence annotation: PelB
sequence: https://synbiohub.org/public/igem/BBa_K2020004_sequence/1

AGC1182TAG
type: SequenceAnnotation
persistent identity: annotation2488234
display id: annotation2488234
version: 1
top level: subE_S221X
role: SO:0001059
role: feature/mutation
location: https://synbiohub.org/public/igem/BBa_K2020004/annotation2488234/range2488234/1

https://synbiohub.org/public/igem/BBa_K2020004/annotation2488234/range2488234/1
type: Range
persistent identity: range2488234
display id: range2488234
version: 1
direction: direction/forward
top level: subE_S221X
start: 1182
end: 1184
orientation: inline

propeptide
type: SequenceAnnotation
persistent identity: annotation2488232
display id: annotation2488232
version: 1
top level: subE_S221X
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K2020004/annotation2488232/range2488232/1

https://synbiohub.org/public/igem/BBa_K2020004/annotation2488232/range2488232/1
type: Range
persistent identity: range2488232
display id: range2488232
version: 1
direction: direction/forward
top level: subE_S221X
start: 293
end: 522
orientation: inline

subtilisin E
type: SequenceAnnotation
persistent identity: annotation2488233
display id: annotation2488233
version: 1
top level: subE_S221X
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K2020004/annotation2488233/range2488233/1

https://synbiohub.org/public/igem/BBa_K2020004/annotation2488233/range2488233/1
type: Range
persistent identity: range2488233
display id: range2488233
version: 1
direction: direction/forward
top level: subE_S221X
start: 523
end: 1351
orientation: inline

BBa_B0010
type: SequenceAnnotation
persistent identity: annotation2488235
display id: annotation2488235
version: 1
top level: subE_S221X
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K2020004/annotation2488235/range2488235/1

https://synbiohub.org/public/igem/BBa_K2020004/annotation2488235/range2488235/1
type: Range
persistent identity: range2488235
display id: range2488235
version: 1
direction: direction/forward
top level: subE_S221X
start: 1360
end: 1439
orientation: inline

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2488230
display id: annotation2488230
version: 1
top level: subE_S221X
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K2020004/annotation2488230/range2488230/1

https://synbiohub.org/public/igem/BBa_K2020004/annotation2488230/range2488230/1
type: Range
persistent identity: range2488230
display id: range2488230
version: 1
direction: direction/forward
top level: subE_S221X
start: 209
end: 220
orientation: inline

LacI
type: SequenceAnnotation
persistent identity: annotation2488229
display id: annotation2488229
version: 1
top level: subE_S221X
role: SO:0000167
role: feature/promoter
location: https://synbiohub.org/public/igem/BBa_K2020004/annotation2488229/range2488229/1

https://synbiohub.org/public/igem/BBa_K2020004/annotation2488229/range2488229/1
type: Range
persistent identity: range2488229
display id: range2488229
version: 1
direction: direction/forward
top level: subE_S221X
start: 1
end: 200
orientation: inline

PelB
type: SequenceAnnotation
persistent identity: annotation2488231
display id: annotation2488231
version: 1
top level: subE_S221X
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K2020004/annotation2488231/range2488231/1

https://synbiohub.org/public/igem/BBa_K2020004/annotation2488231/range2488231/1
type: Range
persistent identity: range2488231
display id: range2488231
version: 1
direction: direction/forward
top level: subE_S221X
start: 227
end: 291
orientation: inline

BBa_K772001
type: ComponentDefinition
persistent identity: BBa K772001
display id: BBa_K772001
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: EpCAM Binding Dock: C-terminus
creator: Mustafa Elitok
created: 2012-09-23T11:00:00Z
modified: 2015-05-08T01:13:15Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _1024_
m user id: 0
owner id: 9573
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: true
mutable description: This gene sequence includes an EpCAM (Epithelial Cell Adhesion Molecule) antigen binding domain, OmpA transmembrane protein (dock) and TEV protease C terminus. It is planned to use this subpart with BBa_772, which contains only TEV protease N terminus differently instead of C terminus, in order to induce a signal reaction in the case of EpCAM presence. EpCAM is an natural antigen which is found high significantly in the cases of some breast and colorectal cancer subtypes. On the upstream region of protein sequence, there are a constitutive pLac promoter and appropiate RBS. To see how this part works: Experience.
mutable notes: There are two protein sequences between domains and the dock: One between the binding domain (antibody) and dock and one between the part of TEV protease.
mutable provenance: CD225 (EpCAM antibody) is commonly used for the test of presence of EpCAM in the laboratories. The sequences of OmpA and TEV protease terminuses are acquired from the Registry.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Translational Unit
role: SO:0000804
sequence annotation: TEV Protease C terminus
sequence annotation: OmpA (Dock)
sequence annotation: RBS
sequence annotation: pCons Promoter
sequence annotation: Binding Dock
sequence annotation: CD215 Antibody
sequence: https://synbiohub.org/public/igem/BBa_K772001_sequence/1

TEV Protease C terminus
type: SequenceAnnotation
persistent identity: annotation2203672
display id: annotation2203672
version: 1
top level: BBa_K772001
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K772001/annotation2203672/range2203672/1

https://synbiohub.org/public/igem/BBa_K772001/annotation2203672/range2203672/1
type: Range
persistent identity: range2203672
display id: range2203672
version: 1
direction: direction/forward
top level: BBa_K772001
start: 94
end: 419
orientation: inline

OmpA (Dock)
type: SequenceAnnotation
persistent identity: annotation2203671
display id: annotation2203671
version: 1
top level: BBa_K772001
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K772001/annotation2203671/range2203671/1

https://synbiohub.org/public/igem/BBa_K772001/annotation2203671/range2203671/1
type: Range
persistent identity: range2203671
display id: range2203671
version: 1
direction: direction/forward
top level: BBa_K772001
start: 420
end: 857
orientation: inline

RBS
type: SequenceAnnotation
persistent identity: annotation2203670
display id: annotation2203670
version: 1
top level: BBa_K772001
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K772001/annotation2203670/range2203670/1

https://synbiohub.org/public/igem/BBa_K772001/annotation2203670/range2203670/1
type: Range
persistent identity: range2203670
display id: range2203670
version: 1
direction: direction/forward
top level: BBa_K772001
start: 73
end: 93
orientation: inline

pCons Promoter
type: SequenceAnnotation
persistent identity: annotation2203669
display id: annotation2203669
version: 1
top level: BBa_K772001
role: SO:0000167
role: feature/promoter
location: https://synbiohub.org/public/igem/BBa_K772001/annotation2203669/range2203669/1

https://synbiohub.org/public/igem/BBa_K772001/annotation2203669/range2203669/1
type: Range
persistent identity: range2203669
display id: range2203669
version: 1
direction: direction/forward
top level: BBa_K772001
start: 1
end: 72
orientation: inline

Binding Dock
type: SequenceAnnotation
persistent identity: annotation2203674
display id: annotation2203674
version: 1
top level: BBa_K772001
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K772001/annotation2203674/range2203674/1

https://synbiohub.org/public/igem/BBa_K772001/annotation2203674/range2203674/1
type: Range
persistent identity: range2203674
display id: range2203674
version: 1
direction: direction/forward
top level: BBa_K772001
start: 94
end: 1159
orientation: inline

CD215 Antibody
type: SequenceAnnotation
persistent identity: annotation2203673
display id: annotation2203673
version: 1
top level: BBa_K772001
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K772001/annotation2203673/range2203673/1

https://synbiohub.org/public/igem/BBa_K772001/annotation2203673/range2203673/1
type: Range
persistent identity: range2203673
display id: range2203673
version: 1
direction: direction/forward
top level: BBa_K772001
start: 858
end: 1159
orientation: inline

subE_Y77W
type: ComponentDefinition
persistent identity: BBa K2020005
display id: BBa_K2020005
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: mutated expression system for subtilisin E in E. coli (Y77W)
creator: Nicola Freyer, Lea Steinbeck, Svenja Meyer, Alexander Deitert
created: 2016-10-07T11:00:00Z
modified: 2016-10-08T12:13:18Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2487_
m user id: 30763
owner id: 30763
owning group id: 9
status: status/Planning
bookmark: false
mutable description: This composite part consists of the promoter BBa_R0010, the ribosome binding site BBa_B0034, the newly created BioBrick part BBa_K2020001 and the terminator BBa_B0010. BioBrick BBa_K2020001 is a composite part itself and includes the secretion tag pelB (BBa_J32015) and a subtilisin E gene optimized for ''Escherichia coli'' codon usage (BBa_K2020000). Once introduced into ''E. coli'', this BioBrick is able to produce an inactive version of subtilisin E and simultaneously secret the enzyme into the periplasm of the cell. Subtilisin E is an alkaline serine protease which non-specifically digests proteins. By performing a site-directed mutagenesis, tyrosine77 in the propeptide of the enzyme was exchanged against tryptophan. Therefore, the enzyme loses its proteolytic activity. With this composite part, iGEM Aachen 2016 was able to prove that tyrosine77 in the propeptide of subtilisin E is essential for the activity of the enzyme.
mutable notes: The sequence of the subtilisin E gene from ''Bacillus subtilis'' was codon optimized for ''E. coli'' with the DNA and protein sequence analysis software "Geneious" and additionally with the "Codon Optimization Tool" from IDT. The sequence was partly ordered from IDT (BBa_K2020001 + BBa_B0010) and then cloned into BBa_J04500, a protein expression backbone which already carries the LacI promoter BBa_R0010 and the ribosome binding site BBa_B0034. Afterwards, a mutation in the active site of the enzyme was introduced by performing a site-directed mutagenesis. The codon TAT of tyrosine77 was substituted with TGG which codes for tryptophan.
mutable provenance: The sequences of the BioBrick parts were obtained through the Registry. {| class="wikitable" style="text-align:center" |- ! part !! colspan="2"| BioBrick No. |- | LacI promoter || colspan="2"| BBa_R0010 |- | RBS || colspan="2"| BBa_B0034 |- | rowspan="2" | CDS || colspan="2"| BBa_K2020001 |- | BBa_J32015 || BBa_K2020000 |- | terminator || colspan="2"| BBa_B0010 |- |} The construct was ordered at IDT.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Composite
sequence annotation: BBa_B0010
sequence annotation: subtilisin E
sequence annotation: propeptide
sequence annotation: PelB
sequence annotation: LacI
sequence annotation: BBa_B0034
sequence annotation: TAT520TGG
sequence: https://synbiohub.org/public/igem/BBa_K2020005_sequence/1

BBa_B0010
type: SequenceAnnotation
persistent identity: annotation2488242
display id: annotation2488242
version: 1
top level: subE_Y77W
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K2020005/annotation2488242/range2488242/1

https://synbiohub.org/public/igem/BBa_K2020005/annotation2488242/range2488242/1
type: Range
persistent identity: range2488242
display id: range2488242
version: 1
direction: direction/forward
top level: subE_Y77W
start: 1360
end: 1439
orientation: inline

subtilisin E
type: SequenceAnnotation
persistent identity: annotation2488241
display id: annotation2488241
version: 1
top level: subE_Y77W
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K2020005/annotation2488241/range2488241/1

https://synbiohub.org/public/igem/BBa_K2020005/annotation2488241/range2488241/1
type: Range
persistent identity: range2488241
display id: range2488241
version: 1
direction: direction/forward
top level: subE_Y77W
start: 523
end: 1351
orientation: inline

propeptide
type: SequenceAnnotation
persistent identity: annotation2488239
display id: annotation2488239
version: 1
top level: subE_Y77W
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K2020005/annotation2488239/range2488239/1

https://synbiohub.org/public/igem/BBa_K2020005/annotation2488239/range2488239/1
type: Range
persistent identity: range2488239
display id: range2488239
version: 1
direction: direction/forward
top level: subE_Y77W
start: 293
end: 522
orientation: inline

PelB
type: SequenceAnnotation
persistent identity: annotation2488238
display id: annotation2488238
version: 1
top level: subE_Y77W
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K2020005/annotation2488238/range2488238/1

https://synbiohub.org/public/igem/BBa_K2020005/annotation2488238/range2488238/1
type: Range
persistent identity: range2488238
display id: range2488238
version: 1
direction: direction/forward
top level: subE_Y77W
start: 227
end: 292
orientation: inline

LacI
type: SequenceAnnotation
persistent identity: annotation2488236
display id: annotation2488236
version: 1
top level: subE_Y77W
role: SO:0000167
role: feature/promoter
location: https://synbiohub.org/public/igem/BBa_K2020005/annotation2488236/range2488236/1

https://synbiohub.org/public/igem/BBa_K2020005/annotation2488236/range2488236/1
type: Range
persistent identity: range2488236
display id: range2488236
version: 1
direction: direction/forward
top level: subE_Y77W
start: 1
end: 200
orientation: inline

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2488237
display id: annotation2488237
version: 1
top level: subE_Y77W
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K2020005/annotation2488237/range2488237/1

https://synbiohub.org/public/igem/BBa_K2020005/annotation2488237/range2488237/1
type: Range
persistent identity: range2488237
display id: range2488237
version: 1
direction: direction/forward
top level: subE_Y77W
start: 209
end: 220
orientation: inline

TAT520TGG
type: SequenceAnnotation
persistent identity: annotation2488240
display id: annotation2488240
version: 1
top level: subE_Y77W
role: SO:0001059
role: feature/mutation
location: https://synbiohub.org/public/igem/BBa_K2020005/annotation2488240/range2488240/1

https://synbiohub.org/public/igem/BBa_K2020005/annotation2488240/range2488240/1
type: Range
persistent identity: range2488240
display id: range2488240
version: 1
direction: direction/forward
top level: subE_Y77W
start: 520
end: 522
orientation: inline

subE_Y77X
type: ComponentDefinition
persistent identity: BBa K2020006
display id: BBa_K2020006
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: mutated expression system for subtilisin E in E. coli (Y77X)
creator: Nicola Freyer, Lea Steinbeck, Svenja Meyer, Alexander Deitert
created: 2016-10-07T11:00:00Z
modified: 2016-10-08T12:30:28Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2487_
m user id: 30763
owner id: 30763
owning group id: 9
status: status/Planning
bookmark: false
mutable description: This composite part consists of the promoter BBa_R0010, the ribosome binding site BBa_B0034, the newly created BioBrick part BBa_K2020001 and the terminator BBa_B0010. BioBrick BBa_K2020001 is a composite part itself and includes the secretion tag pelB (BBa_J32015) and a subtilisin E gene optimized for ''Escherichia coli'' codon usage (BBa_K2020000). Once introduced into ''E. coli'', this BioBrick is able to produce an inactive version of subtilisin E and simultaneously secret the enzyme into the periplasm of the cell. Subtilisin E is an alkaline serine protease which non-specifically digests proteins. By performing a site-directed mutagenesis, tyrosine77 in the propeptide of the enzyme was exchanged against the amber stop codon UAG. Therefore, the expression of the enzyme is interrupted. This composite part was created to integrate a non-canonical amino acid into the sequence of the propeptide of subtilisin E by adding an orthogonal tRNA/aminoacyl-synthetase pair that is capable of incorporating the non-canonical amino acid of choice at the UAG codon.
mutable notes: The sequence of the subtilisin E gene from ''Bacillus subtilis'' was codon optimized for ''E. coli'' with the DNA and protein sequence analysis software "Geneious" and additionally with the "Codon Optimization Tool" from IDT. The sequence was partly ordered from IDT (BBa_K2020001 + BBa_B0010) and then cloned into BBa_J04500, a protein expression backbone which already carries the LacI promoter BBa_R0010 and the ribosome binding site BBa_B0034. Afterwards, a mutation in the active site of the enzyme was introduced by performing a site-directed mutagenesis. The codon TAT of tyrosine77 was substituted with the stop codon TAG.
mutable provenance: The sequences of the BioBrick parts were obtained through the Registry. {| class="wikitable" style="text-align:center" |- ! part !! colspan="2"| BioBrick No. |- | LacI promoter || colspan="2"| BBa_R0010 |- | RBS || colspan="2"| BBa_B0034 |- | rowspan="2" | CDS || colspan="2"| BBa_K2020001 |- | BBa_J32015 || BBa_K2020000 |- | terminator || colspan="2"| BBa_B0010 |- |} The construct was ordered at IDT.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Composite
sequence annotation: BBa_B0034
sequence annotation: PelB
sequence annotation: LacI
sequence annotation: TGG520TAG
sequence annotation: subtilisin E
sequence annotation: BBa_B0010
sequence annotation: propeptide
sequence: https://synbiohub.org/public/igem/BBa_K2020006_sequence/1

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2488249
display id: annotation2488249
version: 1
top level: subE_Y77X
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K2020006/annotation2488249/range2488249/1

https://synbiohub.org/public/igem/BBa_K2020006/annotation2488249/range2488249/1
type: Range
persistent identity: range2488249
display id: range2488249
version: 1
direction: direction/forward
top level: subE_Y77X
start: 209
end: 220
orientation: inline

PelB
type: SequenceAnnotation
persistent identity: annotation2488250
display id: annotation2488250
version: 1
top level: subE_Y77X
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K2020006/annotation2488250/range2488250/1

https://synbiohub.org/public/igem/BBa_K2020006/annotation2488250/range2488250/1
type: Range
persistent identity: range2488250
display id: range2488250
version: 1
direction: direction/forward
top level: subE_Y77X
start: 227
end: 292
orientation: inline

LacI
type: SequenceAnnotation
persistent identity: annotation2488248
display id: annotation2488248
version: 1
top level: subE_Y77X
role: SO:0000167
role: feature/promoter
location: https://synbiohub.org/public/igem/BBa_K2020006/annotation2488248/range2488248/1

https://synbiohub.org/public/igem/BBa_K2020006/annotation2488248/range2488248/1
type: Range
persistent identity: range2488248
display id: range2488248
version: 1
direction: direction/forward
top level: subE_Y77X
start: 1
end: 200
orientation: inline

TGG520TAG
type: SequenceAnnotation
persistent identity: annotation2488252
display id: annotation2488252
version: 1
top level: subE_Y77X
role: SO:0001059
role: feature/mutation
location: https://synbiohub.org/public/igem/BBa_K2020006/annotation2488252/range2488252/1

https://synbiohub.org/public/igem/BBa_K2020006/annotation2488252/range2488252/1
type: Range
persistent identity: range2488252
display id: range2488252
version: 1
direction: direction/forward
top level: subE_Y77X
start: 520
end: 522
orientation: inline

subtilisin E
type: SequenceAnnotation
persistent identity: annotation2488255
display id: annotation2488255
version: 1
top level: subE_Y77X
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K2020006/annotation2488255/range2488255/1

https://synbiohub.org/public/igem/BBa_K2020006/annotation2488255/range2488255/1
type: Range
persistent identity: range2488255
display id: range2488255
version: 1
direction: direction/forward
top level: subE_Y77X
start: 523
end: 1351
orientation: inline

BBa_B0010
type: SequenceAnnotation
persistent identity: annotation2488257
display id: annotation2488257
version: 1
top level: subE_Y77X
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K2020006/annotation2488257/range2488257/1

https://synbiohub.org/public/igem/BBa_K2020006/annotation2488257/range2488257/1
type: Range
persistent identity: range2488257
display id: range2488257
version: 1
direction: direction/forward
top level: subE_Y77X
start: 1360
end: 1439
orientation: inline

propeptide
type: SequenceAnnotation
persistent identity: annotation2488251
display id: annotation2488251
version: 1
top level: subE_Y77X
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K2020006/annotation2488251/range2488251/1

https://synbiohub.org/public/igem/BBa_K2020006/annotation2488251/range2488251/1
type: Range
persistent identity: range2488251
display id: range2488251
version: 1
direction: direction/forward
top level: subE_Y77X
start: 293
end: 522
orientation: inline

BBa_K1639008
type: ComponentDefinition
persistent identity: BBa K1639008
display id: BBa_K1639008
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Tev Protease, single S219V mutation in the internal cleavage site
creator: Mustafa Yılmaz
created: 2015-09-12T11:00:00Z
modified: 2015-09-18T07:12:27Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _2056_
m user id: 24789
owner id: 20835
owning group id: 9
status: status/Planning
bookmark: false
mutable description: TEV protease (also called Tobacco Etch Virus nuclear inclusion a endopeptidase) is a highly sequence-specific cysteine protease from Tobacco Etch Virus (TEV).The consensus for these native cut sites is ENLYFQ\S where ???\??? denotes the cleaved peptide bond. One of the main uses of this protein is for removing affinity tags from purified proteins. The reason for the use of TEV protease as a biochemical tool is its high sequence specificity. This specificity allows for the controlled cleavage of proteins when the preference sequence is inserted into flexible loops. It also makes it relatively non-toxic in vivo as the recognized sequence scarcely occurs in proteins.[1]
mutable notes: This part contains an additional BamHI restriction site at beginning of Tev Protease sequence and a XhoI restriction site at the end.
mutable provenance: Tobacco etch virus (TEV).
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: ATG Start Codon
sequence annotation: S219V
sequence annotation: Tev Protease
sequence annotation: Stop Codon
sequence: https://synbiohub.org/public/igem/BBa_K1639008_sequence/1

ATG Start Codon
type: SequenceAnnotation
persistent identity: annotation2453252
display id: annotation2453252
version: 1
top level: BBa_K1639008
role: SO:0000318
role: feature/start
location: https://synbiohub.org/public/igem/BBa_K1639008/annotation2453252/range2453252/1

https://synbiohub.org/public/igem/BBa_K1639008/annotation2453252/range2453252/1
type: Range
persistent identity: range2453252
display id: range2453252
version: 1
direction: direction/forward
top level: BBa_K1639008
start: 9
end: 11
orientation: inline

S219V
type: SequenceAnnotation
persistent identity: annotation2453264
display id: annotation2453264
version: 1
top level: BBa_K1639008
role: SO:0001059
role: feature/mutation
location: https://synbiohub.org/public/igem/BBa_K1639008/annotation2453264/range2453264/1

https://synbiohub.org/public/igem/BBa_K1639008/annotation2453264/range2453264/1
type: Range
persistent identity: range2453264
display id: range2453264
version: 1
direction: direction/forward
top level: BBa_K1639008
start: 666
end: 668
orientation: inline

Tev Protease
type: SequenceAnnotation
persistent identity: annotation2453253
display id: annotation2453253
version: 1
top level: BBa_K1639008
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1639008/annotation2453253/range2453253/1

https://synbiohub.org/public/igem/BBa_K1639008/annotation2453253/range2453253/1
type: Range
persistent identity: range2453253
display id: range2453253
version: 1
direction: direction/forward
top level: BBa_K1639008
start: 12
end: 737
orientation: inline

Stop Codon
type: SequenceAnnotation
persistent identity: annotation2453254
display id: annotation2453254
version: 1
top level: BBa_K1639008
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K1639008/annotation2453254/range2453254/1

https://synbiohub.org/public/igem/BBa_K1639008/annotation2453254/range2453254/1
type: Range
persistent identity: range2453254
display id: range2453254
version: 1
direction: direction/forward
top level: BBa_K1639008
start: 738
end: 740
orientation: inline

BBa_K1378001
type: ComponentDefinition
persistent identity: BBa K1378001
display id: BBa_K1378001
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: MlrA
creator: Jin Yuji
created: 2014-09-11T11:00:00Z
modified: 2015-05-08T01:10:12Z
discontinued: false
dominant: false
experience: experience/None
group u list: _1755_
m user id: 0
owner id: 21439
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: true
mutable description: This part does not exist
mutable notes: No
mutable provenance: No
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: cds
sequence: https://synbiohub.org/public/igem/BBa_K1378001_sequence/1

cds
type: SequenceAnnotation
persistent identity: annotation2387419
display id: annotation2387419
version: 1
top level: BBa_K1378001
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1378001/annotation2387419/range2387419/1

https://synbiohub.org/public/igem/BBa_K1378001/annotation2387419/range2387419/1
type: Range
persistent identity: range2387419
display id: range2387419
version: 1
direction: direction/forward
top level: BBa_K1378001
start: 1
end: 1011
orientation: inline

BBa_K1319004
type: ComponentDefinition
persistent identity: BBa K1319004
display id: BBa_K1319004
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: TEV protease with anti-self cleavage mutation S219V, codon optimized for E. coli
creator: Florian Gohr, Michael Osthege
created: 2014-09-24T11:00:00Z
modified: 2015-05-08T01:09:50Z
discontinued: false
dominant: false
experience: experience/None
group u list: _1694_
m user id: 0
owner id: 19522
owning group id: 9
sample status: In stock
status: status/Available
bookmark: false
mutable description: This part is a TEV protease in RFC25 that was optimized for expression in E. coli. The part contains the S219V anti-self cleavage mutation.
mutable notes: The part does not contain start or stop codons, because it was designed for RFC25 compability
mutable provenance: The part was codon optimized using the genomic amino acid sequence, designed for RFC25 and mutated S219V for anti-self cleavage properties.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: S219V
sequence annotation: TEV protease
sequence: https://synbiohub.org/public/igem/BBa_K1319004_sequence/1

S219V
type: SequenceAnnotation
persistent identity: annotation2386540
display id: annotation2386540
version: 1
top level: BBa_K1319004
role: SO:0001059
role: feature/mutation
location: https://synbiohub.org/public/igem/BBa_K1319004/annotation2386540/range2386540/1

https://synbiohub.org/public/igem/BBa_K1319004/annotation2386540/range2386540/1
type: Range
persistent identity: range2386540
display id: range2386540
version: 1
direction: direction/forward
top level: BBa_K1319004
start: 655
end: 657
orientation: inline

TEV protease
type: SequenceAnnotation
persistent identity: annotation2386539
display id: annotation2386539
version: 1
top level: BBa_K1319004
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1319004/annotation2386539/range2386539/1

https://synbiohub.org/public/igem/BBa_K1319004/annotation2386539/range2386539/1
type: Range
persistent identity: range2386539
display id: range2386539
version: 1
direction: direction/forward
top level: BBa_K1319004
start: 1
end: 726
orientation: inline

BBa_I712020
type: ComponentDefinition
persistent identity: BBa I712020
display id: BBa_I712020
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Homo sapiens caspase 3
creator: Anja Korenčič
created: 2007-10-20T11:00:00Z
modified: 2015-08-31T04:07:46Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _130_
m user id: 0
owner id: 1846
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable description: Homo sapiens caspase 3, apoptosis-related cysteine peptidase (transcript variant beta).
mutable notes: Includes start and stop codon. The part is cloned into BBa_J52017, a vector with eukaryotic terminator.
mutable provenance: Part comes from Origene SC101958 vector (NM_032991).
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence: https://synbiohub.org/public/igem/BBa_I712020_sequence/1