BBa_K316009
type: ComponentDefinition
persistent identity: BBa K316009
display id: BBa_K316009
version: 1
was generated by: iGEM to SBOL conversion
description: LacI inducible Fast Response Module, using cleavable XylE
created: 2010-10-24T11:00:00Z
modified: 2015-05-08T01:11:56Z
mutable provenance: Made from parts - <bbpart>BBa_k316012</bbpart> and <bbpart>BBa_k316008</bbpart>
owned by: james
owned by: myers
mutable description: 5' his tagged GFP-TEV linker-XylE construct is pre-made in the cell under constitutive promoter. Both Pveg promoter and spoVG RBS are best suited for maximal expression in B.subtilis. While GFP is attached to the XylE monomer via the TEV cleavable linker, the catalytic activity is low. Transcription of TEV protease allows cleavage of the linker between GFP and XylE, thus XylE is free to tetramerise into a fully functional enzyme. XylE is then able to act as described in <bbpart>BBa_k316004</bbpart> Please see 'Part Design' section for design considerations and parts used.
discontinued: false
dominant: false
experience: experience/None
group u list: _440_
m user id: 0
owner id: 7480
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable notes: Standard biobrick assembly [http://partsregistry.org/Assembly:Standard_assembly]
star: false
creator: IC 2010 Team
type: DnaRegion
role: SO:0000804
role: partType/Composite
component: BBa_B0014
component: BBa_K316006
component: BBa_K143053
component: BBa_K316017
sequence annotation: BBa_K316017
sequence annotation: BBa_K143053
sequence annotation: BBa_K316006
sequence annotation: BBa_B0014
sequence: https://synbiohub.org/public/igem/BBa_K316009_sequence/1

BBa_B0014
type: Component
persistent identity: component2225306
display id: component2225306
version: 1
top level: BBa_K316009
definition: BBa_B0014
access: public

BBa_K316006
type: Component
persistent identity: component2225299
display id: component2225299
version: 1
top level: BBa_K316009
definition: GFP-XylE
access: public

BBa_K143053
type: Component
persistent identity: component2225291
display id: component2225291
version: 1
top level: BBa_K316009
definition: Pveg-spoVG
access: public

BBa_K316017
type: Component
persistent identity: component2225285
display id: component2225285
version: 1
top level: BBa_K316009
definition: BBa_K316017
access: public

BBa_K316017
type: SequenceAnnotation
persistent identity: annotation2225285
display id: annotation2225285
version: 1
top level: BBa_K316009
location: https://synbiohub.org/public/igem/BBa_K316009/annotation2225285/range2225285/1
component: BBa_K316017

https://synbiohub.org/public/igem/BBa_K316009/annotation2225285/range2225285/1
type: Range
persistent identity: range2225285
display id: range2225285
version: 1
top level: BBa_K316009
direction: direction/reverse
start: 1
end: 874
orientation: inline

BBa_K143053
type: SequenceAnnotation
persistent identity: annotation2225291
display id: annotation2225291
version: 1
top level: BBa_K316009
location: https://synbiohub.org/public/igem/BBa_K316009/annotation2225291/range2225291/1
component: BBa_K143053

https://synbiohub.org/public/igem/BBa_K316009/annotation2225291/range2225291/1
type: Range
persistent identity: range2225291
display id: range2225291
version: 1
top level: BBa_K316009
direction: direction/reverse
start: 883
end: 999
orientation: inline

BBa_K316006
type: SequenceAnnotation
persistent identity: annotation2225299
display id: annotation2225299
version: 1
top level: BBa_K316009
location: https://synbiohub.org/public/igem/BBa_K316009/annotation2225299/range2225299/1
component: BBa_K316006

https://synbiohub.org/public/igem/BBa_K316009/annotation2225299/range2225299/1
type: Range
persistent identity: range2225299
display id: range2225299
version: 1
top level: BBa_K316009
direction: direction/reverse
start: 1006
end: 2727
orientation: inline

BBa_B0014
type: SequenceAnnotation
persistent identity: annotation2225306
display id: annotation2225306
version: 1
top level: BBa_K316009
location: https://synbiohub.org/public/igem/BBa_K316009/annotation2225306/range2225306/1
component: BBa_B0014

https://synbiohub.org/public/igem/BBa_K316009/annotation2225306/range2225306/1
type: Range
persistent identity: range2225306
display id: range2225306
version: 1
top level: BBa_K316009
direction: direction/reverse
start: 2736
end: 2830
orientation: inline

BBa_B0014
type: ComponentDefinition
persistent identity: BBa B0014
display id: BBa_B0014
version: 1
was generated by: iGEM to SBOL conversion
description: double terminator (B0012-B0011)
created: 2003-07-15T11:00:00Z
modified: 2015-08-31T04:07:20Z
owned by: james
owned by: myers
part status: Released HQ 2013
mutable description: Double terminator consisting of BBa_B0012 and BBa_B0011
discontinued: false
dominant: true
experience: experience/Works
group u list: _1_
m user id: 0
owner id: 24
owning group id: 7
sample status: In stock
status: status/Available
bookmark: false
star: true
creator: Reshma Shetty
type: DnaRegion
role: SO:0000141
role: partType/Terminator
component: BBa_B0011
component: BBa_B0012
sequence annotation: BBa_B0012
sequence annotation: BBa_B0011
sequence: https://synbiohub.org/public/igem/BBa_B0014_sequence/1

BBa_B0011
type: Component
persistent identity: component939311
display id: component939311
version: 1
top level: BBa_B0014
definition: BBa_B0011
access: public

BBa_B0012
type: Component
persistent identity: component939303
display id: component939303
version: 1
top level: BBa_B0014
definition: BBa_B0012
access: public

BBa_B0012
type: SequenceAnnotation
persistent identity: annotation939303
display id: annotation939303
version: 1
top level: BBa_B0014
location: https://synbiohub.org/public/igem/BBa_B0014/annotation939303/range939303/1
component: BBa_B0012

https://synbiohub.org/public/igem/BBa_B0014/annotation939303/range939303/1
type: Range
persistent identity: range939303
display id: range939303
version: 1
top level: BBa_B0014
direction: direction/forward
start: 1
end: 41
orientation: inline

BBa_B0011
type: SequenceAnnotation
persistent identity: annotation939311
display id: annotation939311
version: 1
top level: BBa_B0014
location: https://synbiohub.org/public/igem/BBa_B0014/annotation939311/range939311/1
component: BBa_B0011

https://synbiohub.org/public/igem/BBa_B0014/annotation939311/range939311/1
type: Range
persistent identity: range939311
display id: range939311
version: 1
top level: BBa_B0014
direction: direction/forward
start: 50
end: 95
orientation: inline

Pveg-spoVG
type: ComponentDefinition
persistent identity: BBa K143053
display id: BBa_K143053
version: 1
was generated by: iGEM to SBOL conversion
description: Promoter Pveg and RBS spoVG for B. subtilis
created: 2008-10-07T11:00:00Z
modified: 2015-05-08T01:10:24Z
mutable provenance: Pveg-spoVG was synthesised by GeneArt
owned by: james
owned by: myers
part status: Released HQ 2013
mutable description: Constitutive promoter veg(<bbpart>BBa_K143012</bbpart>) coupled to the strong Ribosome Binding Site spoVG(<bbpart>BBa_K143021</bbpart>) from ''B. subtilis''. Pveg-spoVG can be used in the context of a '''Ribosomes per second''' (RiPS) output generator '''To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix1. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.'''
discontinued: false
dominant: true
experience: experience/Works
group u list: _199_
m user id: 0
owner id: 3475
owning group id: 9
sample status: In stock
status: status/Available
bookmark: false
mutable notes: The sequence of Pveg was obtained from the DBTBS1 and RBS-spoVG were obtained from papers2 and the sequence synthesised by GeneArt
star: true
creator: Chris Hirst
type: DnaRegion
role: SO:0000804
role: partType/Generator
component: BBa_K143012
component: BBa_K143021
sequence annotation: BBa_K143012
sequence annotation: BBa_K143021
sequence: https://synbiohub.org/public/igem/BBa_K143053_sequence/1

BBa_K143012
type: Component
persistent identity: component1979395
display id: component1979395
version: 1
top level: Pveg-spoVG
definition: Pveg
access: public

BBa_K143021
type: Component
persistent identity: component1979397
display id: component1979397
version: 1
top level: Pveg-spoVG
definition: RBS-spoVG
access: public

BBa_K143012
type: SequenceAnnotation
persistent identity: annotation1979395
display id: annotation1979395
version: 1
top level: Pveg-spoVG
location: https://synbiohub.org/public/igem/BBa_K143053/annotation1979395/range1979395/1
component: BBa_K143012

https://synbiohub.org/public/igem/BBa_K143053/annotation1979395/range1979395/1
type: Range
persistent identity: range1979395
display id: range1979395
version: 1
top level: Pveg-spoVG
direction: direction/reverse
start: 1
end: 97
orientation: inline

BBa_K143021
type: SequenceAnnotation
persistent identity: annotation1979397
display id: annotation1979397
version: 1
top level: Pveg-spoVG
location: https://synbiohub.org/public/igem/BBa_K143053/annotation1979397/range1979397/1
component: BBa_K143021

https://synbiohub.org/public/igem/BBa_K143053/annotation1979397/range1979397/1
type: Range
persistent identity: range1979397
display id: range1979397
version: 1
top level: Pveg-spoVG
direction: direction/reverse
start: 106
end: 117
orientation: inline

GFP-XylE
type: ComponentDefinition
persistent identity: BBa K316006
display id: BBa_K316006
version: 1
was generated by: iGEM to SBOL conversion
description: N-terminus his tagged-GFP-XylE fusion protein
created: 2010-10-20T11:00:00Z
modified: 2015-05-08T01:11:56Z
mutable provenance: Existing biobrick parts with modifications using PCR primer extension
owned by: james
owned by: myers
mutable description: Constructed to be combined with promoter and terminator. The GFP is linked to XylE monomer subunit by a GGGSGGGS linker with the aim to render the enzyme inactive, via preventing tetramerization (it???s functional form).
discontinued: false
dominant: false
experience: experience/Works
group u list: _440_
m user id: 0
owner id: 7480
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable notes: Standard biobrick assembly and PCR primer extension. For full methods please see our wiki http://2010.igem.org/Team:Imperial_College_London/Strategy
star: false
creator: IC 2010 Team
type: DnaRegion
role: partType/Composite
role: SO:0000804
sequence annotation: Start
sequence annotation: His tag
sequence annotation: GFP E0040
sequence annotation: Flag Tag
sequence annotation: Tev Protease Cleavage Site
sequence annotation: GGS linker
sequence annotation: XylE J33204
sequence: https://synbiohub.org/public/igem/BBa_K316006_sequence/1

Start
type: SequenceAnnotation
persistent identity: annotation2094917
display id: annotation2094917
version: 1
top level: GFP-XylE
location: https://synbiohub.org/public/igem/BBa_K316006/annotation2094917/range2094917/1
role: feature/start
role: SO:0000318

https://synbiohub.org/public/igem/BBa_K316006/annotation2094917/range2094917/1
type: Range
persistent identity: range2094917
display id: range2094917
version: 1
top level: GFP-XylE
direction: direction/forward
start: 1
end: 3
orientation: inline

His tag
type: SequenceAnnotation
persistent identity: annotation2094916
display id: annotation2094916
version: 1
top level: GFP-XylE
location: https://synbiohub.org/public/igem/BBa_K316006/annotation2094916/range2094916/1
role: SO:0000324
role: feature/tag

https://synbiohub.org/public/igem/BBa_K316006/annotation2094916/range2094916/1
type: Range
persistent identity: range2094916
display id: range2094916
version: 1
top level: GFP-XylE
direction: direction/forward
start: 4
end: 21
orientation: inline

GFP E0040
type: SequenceAnnotation
persistent identity: annotation2094918
display id: annotation2094918
version: 1
top level: GFP-XylE
location: https://synbiohub.org/public/igem/BBa_K316006/annotation2094918/range2094918/1
role: SO:0000316
role: feature/protein

https://synbiohub.org/public/igem/BBa_K316006/annotation2094918/range2094918/1
type: Range
persistent identity: range2094918
display id: range2094918
version: 1
top level: GFP-XylE
direction: direction/forward
start: 22
end: 732
orientation: inline

Flag Tag
type: SequenceAnnotation
persistent identity: annotation2094919
display id: annotation2094919
version: 1
top level: GFP-XylE
location: https://synbiohub.org/public/igem/BBa_K316006/annotation2094919/range2094919/1
role: SO:0000324
role: feature/tag

https://synbiohub.org/public/igem/BBa_K316006/annotation2094919/range2094919/1
type: Range
persistent identity: range2094919
display id: range2094919
version: 1
top level: GFP-XylE
direction: direction/forward
start: 733
end: 756
orientation: inline

Tev Protease Cleavage Site
type: SequenceAnnotation
persistent identity: annotation2094920
display id: annotation2094920
version: 1
top level: GFP-XylE
location: https://synbiohub.org/public/igem/BBa_K316006/annotation2094920/range2094920/1
role: feature/misc
role: SO:0000110

https://synbiohub.org/public/igem/BBa_K316006/annotation2094920/range2094920/1
type: Range
persistent identity: range2094920
display id: range2094920
version: 1
top level: GFP-XylE
direction: direction/forward
start: 757
end: 777
orientation: inline

GGS linker
type: SequenceAnnotation
persistent identity: annotation2094921
display id: annotation2094921
version: 1
top level: GFP-XylE
location: https://synbiohub.org/public/igem/BBa_K316006/annotation2094921/range2094921/1
role: feature/misc
role: SO:0000110

https://synbiohub.org/public/igem/BBa_K316006/annotation2094921/range2094921/1
type: Range
persistent identity: range2094921
display id: range2094921
version: 1
top level: GFP-XylE
direction: direction/forward
start: 778
end: 795
orientation: inline

XylE J33204
type: SequenceAnnotation
persistent identity: annotation2094922
display id: annotation2094922
version: 1
top level: GFP-XylE
location: https://synbiohub.org/public/igem/BBa_K316006/annotation2094922/range2094922/1
role: SO:0000316
role: feature/protein

https://synbiohub.org/public/igem/BBa_K316006/annotation2094922/range2094922/1
type: Range
persistent identity: range2094922
display id: range2094922
version: 1
top level: GFP-XylE
direction: direction/forward
start: 796
end: 1722
orientation: inline

BBa_K316017
type: ComponentDefinition
persistent identity: BBa K316017
display id: BBa_K316017
version: 1
was generated by: iGEM to SBOL conversion
description: LacI operator controlled TEV protease
created: 2010-10-24T11:00:00Z
modified: 2015-05-08T01:11:56Z
mutable provenance: The part was produced by nucleotide synthesis by mwg ??? eurofins. The synthetic RBS was designed to work with <bbpart>BBa_K316000</bbpart>.
owned by: james
owned by: myers
mutable description: TEV protease S219P autocatalysis resistant variant <bbpart>BBa_K316012</bbpart>. This part had been reversed for the 3' strand in order to reduce any read-through that may be caused by upstream elements. Expression of TEV protease is under control of enhanced hyperspank promoter <bbpart>BBa_K316000</bbpart>.
discontinued: false
dominant: false
experience: experience/None
group u list: _440_
m user id: 0
owner id: 7480
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable notes: TEV protease is naturally found in Tobacco Etch Virus genome. This part contains the sequence codon optimized for expression in B.subtilis using mwg ??? eurofins [http://www.eurofinsdna.com/home.html] proprietary software.
star: false
creator: IC 2010 Team
type: DnaRegion
role: SO:0000316
role: partType/Coding
component: BBa_K316000
component: BBa_K316012
sequence annotation: BBa_K316012
sequence annotation: BBa_K316000
sequence: https://synbiohub.org/public/igem/BBa_K316017_sequence/1

BBa_K316000
type: Component
persistent identity: component2218950
display id: component2218950
version: 1
top level: BBa_K316017
definition: sRBS Pehs
access: public

BBa_K316012
type: Component
persistent identity: component2218945
display id: component2218945
version: 1
top level: BBa_K316017
definition: TEV
access: public

BBa_K316012
type: SequenceAnnotation
persistent identity: annotation2218945
display id: annotation2218945
version: 1
top level: BBa_K316017
location: https://synbiohub.org/public/igem/BBa_K316017/annotation2218945/range2218945/1
component: BBa_K316012

https://synbiohub.org/public/igem/BBa_K316017/annotation2218945/range2218945/1
type: Range
persistent identity: range2218945
display id: range2218945
version: 1
top level: BBa_K316017
direction: direction/reverse
start: 1
end: 717
orientation: inline

BBa_K316000
type: SequenceAnnotation
persistent identity: annotation2218950
display id: annotation2218950
version: 1
top level: BBa_K316017
location: https://synbiohub.org/public/igem/BBa_K316017/annotation2218950/range2218950/1
component: BBa_K316000

https://synbiohub.org/public/igem/BBa_K316017/annotation2218950/range2218950/1
type: Range
persistent identity: range2218950
display id: range2218950
version: 1
top level: BBa_K316017
direction: direction/reverse
start: 726
end: 874
orientation: inline

BBa_B0011
type: ComponentDefinition
persistent identity: BBa B0011
display id: BBa_B0011
version: 1
was generated by: iGEM to SBOL conversion
description: LuxICDABEG (+/-)
created: 2003-01-31T12:00:00Z
modified: 2015-08-31T04:07:20Z
mutable provenance: Derived from luxICDABEG operon terminator of Vibrio fischeri <genbank>AF170104</genbank>.
owned by: james
owned by: myers
part status: Released HQ 2013
mutable description: Bidirectional transcriptional terminator consisting of a 22 bp stem-loop.
discontinued: false
dominant: false
experience: experience/Works
group u list: _1_
m user id: 0
owner id: 24
owning group id: 7
sample status: In stock
status: status/Available
bookmark: false
mutable notes: In the naturally-occuring sequence there is a mismatch in the stem of the stem loop. This can be corrected via an A->G mutation (at position 40 -- sequence coordinate/not MFOLD coordinate). The above sequence does not reflect this mutation (but the MFOLD image does). This terminator's location cannot be found using some inverted repeat detectors like PALINDROME because it is too short and contains a mismatch. This one was found with the help of Tom Knight. It lies between two coding regions that point towards eachother.
star: true
creator: Reshma Shetty
type: DnaRegion
role: SO:0000141
role: partType/Terminator
sequence annotation: BBa_B0011
sequence annotation: stem_loop
sequence: https://synbiohub.org/public/igem/BBa_B0011_sequence/1

BBa_B0011
type: SequenceAnnotation
persistent identity: annotation7019
display id: annotation7019
version: 1
top level: BBa_B0011
location: https://synbiohub.org/public/igem/BBa_B0011/annotation7019/range7019/1
role: SO:0000804
role: feature/BioBrick

https://synbiohub.org/public/igem/BBa_B0011/annotation7019/range7019/1
type: Range
persistent identity: range7019
display id: range7019
version: 1
top level: BBa_B0011
direction: direction/forward
start: 1
end: 46
orientation: inline

stem_loop
type: SequenceAnnotation
persistent identity: annotation1683
display id: annotation1683
version: 1
top level: BBa_B0011
location: https://synbiohub.org/public/igem/BBa_B0011/annotation1683/range1683/1
role: SO:0000313
role: feature/stem loop

https://synbiohub.org/public/igem/BBa_B0011/annotation1683/range1683/1
type: Range
persistent identity: range1683
display id: range1683
version: 1
top level: BBa_B0011
direction: direction/forward
start: 13
end: 35
orientation: inline

BBa_B0012
type: ComponentDefinition
persistent identity: BBa B0012
display id: BBa_B0012
version: 1
was generated by: iGEM to SBOL conversion
description: TE from coliphageT7
created: 2003-01-31T12:00:00Z
modified: 2015-08-31T04:07:20Z
mutable provenance: Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>.
owned by: james
owned by: myers
part status: Released HQ 2013
mutable description: Transcription terminator for the E.coli RNA polymerase.
discontinued: false
dominant: false
experience: experience/Fails
group u list: _1_
m user id: 0
owner id: 24
owning group id: 7
sample status: In stock
status: status/Available
bookmark: false
mutable notes: Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG). Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator.
star: false
creator: Reshma Shetty
type: DnaRegion
role: SO:0000141
role: partType/Terminator
sequence annotation: BBa_B0012
sequence annotation: T7 TE
sequence annotation: polya
sequence annotation: stop
sequence: https://synbiohub.org/public/igem/BBa_B0012_sequence/1

BBa_B0012
type: SequenceAnnotation
persistent identity: annotation7020
display id: annotation7020
version: 1
top level: BBa_B0012
location: https://synbiohub.org/public/igem/BBa_B0012/annotation7020/range7020/1
role: feature/BioBrick
role: SO:0000804

https://synbiohub.org/public/igem/BBa_B0012/annotation7020/range7020/1
type: Range
persistent identity: range7020
display id: range7020
version: 1
top level: BBa_B0012
direction: direction/forward
start: 1
end: 41
orientation: inline

T7 TE
type: SequenceAnnotation
persistent identity: annotation1686
display id: annotation1686
version: 1
top level: BBa_B0012
location: https://synbiohub.org/public/igem/BBa_B0012/annotation1686/range1686/1
role: SO:0000313
role: feature/stem loop

https://synbiohub.org/public/igem/BBa_B0012/annotation1686/range1686/1
type: Range
persistent identity: range1686
display id: range1686
version: 1
top level: BBa_B0012
direction: direction/forward
start: 8
end: 27
orientation: inline

polya
type: SequenceAnnotation
persistent identity: annotation1690
display id: annotation1690
version: 1
top level: BBa_B0012
location: https://synbiohub.org/public/igem/BBa_B0012/annotation1690/range1690/1
role: feature/polya
role: SO:0000553

https://synbiohub.org/public/igem/BBa_B0012/annotation1690/range1690/1
type: Range
persistent identity: range1690
display id: range1690
version: 1
top level: BBa_B0012
direction: direction/forward
start: 28
end: 41
orientation: inline

stop
type: SequenceAnnotation
persistent identity: annotation1687
display id: annotation1687
version: 1
top level: BBa_B0012
location: https://synbiohub.org/public/igem/BBa_B0012/annotation1687/range1687/1
role: feature/stop
role: SO:0000319

https://synbiohub.org/public/igem/BBa_B0012/annotation1687/range1687/1
type: Range
persistent identity: range1687
display id: range1687
version: 1
top level: BBa_B0012
direction: direction/forward
start: 34
end: 34
orientation: inline

Pveg
type: ComponentDefinition
persistent identity: BBa K143012
display id: BBa_K143012
version: 1
was generated by: iGEM to SBOL conversion
description: Promoter veg a constitutive promoter for B. subtilis
created: 2008-09-10T11:00:00Z
modified: 2015-05-08T01:10:23Z
mutable provenance: The Pveg promoter was suggested to us by Dr. Jan-Willem Veening of Newcastle University. This sequence supplied was compared to that of the DBTBS database#3 then a section containing the binding site synthesised by Geneart.
owned by: james
owned by: myers
part status: Released HQ 2013
mutable description: Pveg is a constitutive promoter that constitutively expresses the P43 protein in ''B.subtilis''. Pveg contains binding sites for the ''B.sutbilis'' major sigma factor#1. Pveg in ''B.subtilis'' utilises two binding sites to cause high expression of genes#2, however our Pveg is lacking the upstream site to give a medium level of gene expression. It has been noted that the sporulation master regulatoion factor spoOA interacts with Pveg though it is not known how#3. The context with which we used the promoter Pveg is as a '''Polymerase Per Second''' (PoPS) generator.
discontinued: false
dominant: true
experience: experience/Works
group u list: _199_
m user id: 0
owner id: 2090
owning group id: 9
sample status: In stock
status: status/Available
bookmark: false
mutable notes: The biobrick part was designed to include a single binding site for the ''B.subtilis major sigma factor. In addition the biobrick standard was applied to the promoter Pveg sequence.
star: false
creator: James Chappell
type: DnaRegion
role: partType/Regulatory
role: SO:0000167
sequence annotation: Sigma A-35
sequence annotation: Sigma A -10
sequence: https://synbiohub.org/public/igem/BBa_K143012_sequence/1

Sigma A-35
type: SequenceAnnotation
persistent identity: annotation1975704
display id: annotation1975704
version: 1
top level: Pveg
location: https://synbiohub.org/public/igem/BBa_K143012/annotation1975704/range1975704/1
role: feature/promoter
role: SO:0000167

https://synbiohub.org/public/igem/BBa_K143012/annotation1975704/range1975704/1
type: Range
persistent identity: range1975704
display id: range1975704
version: 1
top level: Pveg
direction: direction/forward
start: 63
end: 68
orientation: inline

Sigma A -10
type: SequenceAnnotation
persistent identity: annotation1975705
display id: annotation1975705
version: 1
top level: Pveg
location: https://synbiohub.org/public/igem/BBa_K143012/annotation1975705/range1975705/1
role: SO:0000167
role: feature/promoter

https://synbiohub.org/public/igem/BBa_K143012/annotation1975705/range1975705/1
type: Range
persistent identity: range1975705
display id: range1975705
version: 1
top level: Pveg
direction: direction/forward
start: 86
end: 91
orientation: inline

RBS-spoVG
type: ComponentDefinition
persistent identity: BBa K143021
display id: BBa_K143021
version: 1
was generated by: iGEM to SBOL conversion
description: SpoVG ribosome binding site (RBS) for B. subtilis
created: 2008-09-16T11:00:00Z
modified: 2015-05-08T01:10:23Z
mutable provenance: The sequence was taken from a previous research paper [1] and was constructed by Geneart.
owned by: james
owned by: myers
part status: Released HQ 2013
mutable description: Description: SpoVG is an endogenous ribosome binding site from B.subtilis. The sequence of the spoVG ribosome binding site is AAAGGUGGUGA which is complementary to the sequence UUUCCUCCACU from the 3' region of the 16s rRNA from B.subtilis. Previous research showed that the predicted binding energy of the 16s rRNA to the RBS is -19kcal 1
discontinued: false
dominant: true
experience: experience/Works
group u list: _199_
m user id: 0
owner id: 2090
owning group id: 9
sample status: In stock
status: status/Available
bookmark: false
mutable notes: In order to ensure that the RBS is functional the actual ribosome binding site was maintained and the distance between the RBS and the start codon maintained. In order to conform to the biobrick standard the sequence flanking the RBS had to be changed but the distance between the promoter and RBS, and start codon and RBS was maintained.
star: false
creator: James Chappell
type: DnaRegion
role: SO:0000139
role: partType/RBS
sequence annotation: rbs
sequence: https://synbiohub.org/public/igem/BBa_K143021_sequence/1

rbs
type: SequenceAnnotation
persistent identity: annotation1975997
display id: annotation1975997
version: 1
top level: RBS-spoVG
location: https://synbiohub.org/public/igem/BBa_K143021/annotation1975997/range1975997/1
role: SO:0000139
role: feature/rbs

https://synbiohub.org/public/igem/BBa_K143021/annotation1975997/range1975997/1
type: Range
persistent identity: range1975997
display id: range1975997
version: 1
top level: RBS-spoVG
direction: direction/forward
start: 1
end: 12
orientation: inline

sRBS Pehs
type: ComponentDefinition
persistent identity: BBa K316000
display id: BBa_K316000
version: 1
was generated by: iGEM to SBOL conversion
description: Reverse strand coding Enhanced LacI-hyperspank promoter
created: 2010-10-19T11:00:00Z
modified: 2015-05-08T01:11:56Z
mutable provenance: DNA synthesis by MWG eurofins. The sequence was codon optimized for expression in B.subtilis using mwg ??? eurofins www.eurofinsdna.com proprietary software.
owned by: james
owned by: myers
mutable description: This part is a modified version of hyper-spank promoter for B.subtilis <bbpart>BBa_K143015</bbpart>. Hyper-spank promoter is repressed by transcriptional repressor LacI <bbpart>BBa_K143033</bbpart> and can be induced by addition of Isopropyl β-D-1-thiogalactopyranoside (IPTG). Constitutive expression of LacI is required for repression. Promoter Design The position and sequence of LacI binding was designed using existing knowledge. The stochastic nature of transcriptional repressors usually leads to background transcription. In order to minimise background the binding sites and the distance between them have been optimised. Stronger binding The natural LacI operator has 3 binding sites, all of which have variations in the binding sequences. Perfectly symmetric binding sequence was shown to have10-fold higher binding compared to wild type sequences. The aattgtgagc gctcacaatt sequence has been shown to be optimal for LacI binding Muller 1996 Oehler 1994. Optimal distance Due to the tetrameric nature of LacI it can simulataneously bind to multiple regions in the genome. Binding at multple sites can produce much stronger repression (muller 1996) by increasing local LacI concentrations. Due to the helical nature of DNA the distance between the operator sites plays an important role in the strength of repression. Maximal repression at 70.5bp, second strongest at 92.5bp and third at 115.5bp
discontinued: false
dominant: false
experience: experience/None
group u list: _440_
m user id: 0
owner id: 7480
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable notes: Designed for minimal basal transcription by altering the LacI binding site sequence and distance between them.
star: false
creator: IC 2010 Team
type: DnaRegion
role: SO:0000167
role: partType/Regulatory
sequence annotation: sRBS
sequence annotation: scar
sequence annotation: LacI binding
sequence: https://synbiohub.org/public/igem/BBa_K316000_sequence/1

sRBS
type: SequenceAnnotation
persistent identity: annotation2097815
display id: annotation2097815
version: 1
top level: sRBS Pehs
location: https://synbiohub.org/public/igem/BBa_K316000/annotation2097815/range2097815/1
role: SO:0000139
role: feature/rbs

https://synbiohub.org/public/igem/BBa_K316000/annotation2097815/range2097815/1
type: Range
persistent identity: range2097815
display id: range2097815
version: 1
top level: sRBS Pehs
direction: direction/reverse
start: 1
end: 12
orientation: inline

scar
type: SequenceAnnotation
persistent identity: annotation2097817
display id: annotation2097817
version: 1
top level: sRBS Pehs
location: https://synbiohub.org/public/igem/BBa_K316000/annotation2097817/range2097817/1
role: SO:0000110
role: feature/misc

https://synbiohub.org/public/igem/BBa_K316000/annotation2097817/range2097817/1
type: Range
persistent identity: range2097817
display id: range2097817
version: 1
top level: sRBS Pehs
direction: direction/reverse
start: 13
end: 20
orientation: inline

LacI binding
type: SequenceAnnotation
persistent identity: annotation2100978
display id: annotation2100978
version: 1
top level: sRBS Pehs
location: https://synbiohub.org/public/igem/BBa_K316000/annotation2100978/range2100978/1
role: feature/binding
role: SO:0001091

https://synbiohub.org/public/igem/BBa_K316000/annotation2100978/range2100978/1
type: Range
persistent identity: range2100978
display id: range2100978
version: 1
top level: sRBS Pehs
direction: direction/reverse
start: 21
end: 51
orientation: inline

LacI binding
type: SequenceAnnotation
persistent identity: annotation2100979
display id: annotation2100979
version: 1
top level: sRBS Pehs
location: https://synbiohub.org/public/igem/BBa_K316000/annotation2100979/range2100979/1
role: SO:0001091
role: feature/binding

https://synbiohub.org/public/igem/BBa_K316000/annotation2100979/range2100979/1
type: Range
persistent identity: range2100979
display id: range2100979
version: 1
top level: sRBS Pehs
direction: direction/reverse
start: 111
end: 140
orientation: inline

TEV
type: ComponentDefinition
persistent identity: BBa K316012
display id: BBa_K316012
version: 1
was generated by: iGEM to SBOL conversion
description: TEV protease S219P autocatalysis resistant variant
created: 2010-10-22T11:00:00Z
modified: 2015-05-08T01:11:56Z
mutable provenance: TEV protease is naturally found in Tobacco Etch Virus genome. This part contains the sequence codon optimized for expression in B.subtilis using mwg ??? eurofins www.eurofinsdna.com proprietary software.
owned by: james
owned by: myers
mutable description: TEV protease S219P autocatalysis resistant variant Introduction : This is the nuclear inclusion protease, endogenous to Tobacco Etch Virus and is used in the late lifecycle to cleave polyprotein precursors. The recognition sequence is ENLYFQG/S [http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6W9V-45PMGK3-9P&_user=217827&_coverDate=02%2F01%2F1994&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_acct=C000011279&_version=1&_urlVersion=0&_userid=217827&md5=e075aad3a349720ad9484095d01a65be&searchtype=a]] between QG or QSDue to it???s stringent sequence specificity, TEV is commonly used to cleave genetically engineered proteins. Uses: TEV proteinase is used to cleave fusion proteins. It is useful due to its high degree of specificity http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6W9V-45PMGK3-9P&_user=217827&_coverDate=02%2F01%2F1994&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_acct=C000011279&_version=1&_urlVersion=0&_userid=217827&md5=e075aad3a349720ad9484095d01a65be&searchtype=a and potential to be used in vivo or in vitro applications. Auto-inactivation Wild type TEV protease also cleaves itself at Met 218 and Ser 219 http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WXR-45R86R6-5X&_user=7635175&_coverDate=06%2F20%2F1995&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_acct=C000011279&_version=1&_urlVersion=0&_userid=7635175&md5=0a1521e783c0bd4126b145f3f6d766d4&searchtype=a . This leads to auto-inactivation of the TEV protease and progressive loss of activity of the protein. The rate of inactivation is proportional to the concentration of protease http://peds.oxfordjournals.org/content/14/12/993.long More stable Mutants have been produced by single amino acid substitutions S219V (AGC(serine) to GTG(valine) and S219P (AGC(serine) to CCG(proline) Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type…
discontinued: false
dominant: false
experience: experience/None
group u list: _440_
m user id: 0
owner id: 7480
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable notes: The part was produced by nucleotide synthesis by mwg ??? eurofin
star: false
creator: IC 2010 Team
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: TEV protease
sequence: https://synbiohub.org/public/igem/BBa_K316012_sequence/1

TEV protease
type: SequenceAnnotation
persistent identity: annotation2100977
display id: annotation2100977
version: 1
top level: TEV
location: https://synbiohub.org/public/igem/BBa_K316012/annotation2100977/range2100977/1
role: SO:0000316
role: feature/protein

https://synbiohub.org/public/igem/BBa_K316012/annotation2100977/range2100977/1
type: Range
persistent identity: range2100977
display id: range2100977
version: 1
top level: TEV
direction: direction/reverse
start: 1
end: 717
orientation: inline

https://synbiohub.org/public/igem/BBa_K316009_sequence/1
type: Sequence
persistent identity: BBa K316009 sequence
display id: BBa_K316009_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: ttattagttcattaactgtgtggcctctttaaccggttgaaacggttcttccggtttatccatgaacactttgtgacctccccaaagaacggaatctgcattcagtctccagcctgaaacccattgttgagcttcttggttcgtcagtaactccatgaagttcttgggaacactcgtaaagtaattattcgtattggtgaagtttgacgcagagtgtatgcccacaataaacccatcccgtgttgacacaagaggagagccacattgcccatcttttgtctgaatccaatgcttccaaaagatgccatcagagctagggaatgtacagctagtgtctgataccattgaggacatagatttggtctgaaagtttgttgtgactaagcagatccgttcttcacgctgaggttcgcgaaacttgagtttctgaggaaacggtggaaagtctttcggcattcgaatgataatcatgtctcttccatctatcagatgttgctgaagcgtcgttgtgtttttgaccttaaagactccatgaagggattgtaccaacagtgtaccattattgcgcctaaagaggtgtttgttcgtgatgatgaatggcccaaagccaatgccatacaagctagtggtatgaccatcactctcattggtcagatggcaaatagtcgagctaatcggattatagtcacgtggtcctttaaacaacgatcttcccattactagagttcacctcctttctctagtatgtgaattgttatccgctcacaattccacacacacattatgccacaccttgtagataaagtcaacaacttttgcaactttctcggcaaaatgtggaattgtgagcgctcacaattccacaaccctcgagtactagagaattttgtcaaaataattttattgacaacgtcttattaacgttgatataatttaaattttatttgacaaaaatgggctcgtgttgtacaataaatgttactagagaaaggtggtgaatactagatgcatcaccatcatcaccatcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaagattataaagatgatgacgacaaggaaaatctttattttcaagggggaggttcaggaggcagcatgaacaaaggtgtaatgcgaccgggccatgtgcagctgcgtgtactggacatgagcaaggccctggaacactacgtcgagttgctgggcctgatcgagatggaccgtgacgaccagggccgtgtctatctgaaggcttggaccgaagtggataagttttccctggtgctacgcgaggctgacgagccgggcatggattt…
encoding: naseq.html

https://synbiohub.org/public/igem/BBa_B0014_sequence/1
type: Sequence
persistent identity: BBa B0014 sequence
display id: BBa_B0014_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: tcacactggctcaccttcgggtgggcctttctgcgtttatatactagagagagaatataaaaagccagattattaatccggcttttttattattt
encoding: naseq.html

https://synbiohub.org/public/igem/BBa_K143053_sequence/1
type: Sequence
persistent identity: BBa K143053 sequence
display id: BBa_K143053_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: aattttgtcaaaataattttattgacaacgtcttattaacgttgatataatttaaattttatttgacaaaaatgggctcgtgttgtacaataaatgttactagagaaaggtggtgaa
encoding: naseq.html

https://synbiohub.org/public/igem/BBa_K316006_sequence/1
type: Sequence
persistent identity: BBa K316006 sequence
display id: BBa_K316006_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: atgcatcaccatcatcaccatcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaagattataaagatgatgacgacaaggaaaatctttattttcaagggggaggttcaggaggcagcatgaacaaaggtgtaatgcgaccgggccatgtgcagctgcgtgtactggacatgagcaaggccctggaacactacgtcgagttgctgggcctgatcgagatggaccgtgacgaccagggccgtgtctatctgaaggcttggaccgaagtggataagttttccctggtgctacgcgaggctgacgagccgggcatggattttatgggtttcaaggttgtggatgaggatgctctccggcaactggagcgggatctgatggcatatggctgtgccgttgagcagctacccgcaggtgaactgaacagttgtggccggcgcgtgcgcttccaggccccctccgggcatcacttcgagttgtatgcagacaaggaatatactggaaagtggggtttgaatgacgtcaatcccgaggcatggccgcgcgatctgaaaggtatggcggctgtgcgtttcgaccacgccctcatgtatggcgacgaattgccggcgacctatgacctgttcaccaaggtgctcggtttctatctggccgaacaggtgctggacgaaaatggcacgcgcgtcgcccagtttctcagtctgtcgaccaaggcccacgacgtggccttcattcaccatccggaaaaaggccgcctccatcatgtgtccttccacctcgaaacctgggaagacttgcttcgcgccgccgacctgatctccatgaccgacacatctatcgatatcggcccaacccgccacggcctcactcacggcaagaccatctacttcttcgacccgtccggtaaccgcaacgaagtgttctgcgggggagattacaactacccggaccacaaaccggtgacctggaccaccgaccagctgggcaaggcgatcttttaccacgaccgcattctcaacgaacgattcatgaccgtgctgacctaataa
encoding: naseq.html

https://synbiohub.org/public/igem/BBa_K316017_sequence/1
type: Sequence
persistent identity: BBa K316017 sequence
display id: BBa_K316017_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: ttattagttcattaactgtgtggcctctttaaccggttgaaacggttcttccggtttatccatgaacactttgtgacctccccaaagaacggaatctgcattcagtctccagcctgaaacccattgttgagcttcttggttcgtcagtaactccatgaagttcttgggaacactcgtaaagtaattattcgtattggtgaagtttgacgcagagtgtatgcccacaataaacccatcccgtgttgacacaagaggagagccacattgcccatcttttgtctgaatccaatgcttccaaaagatgccatcagagctagggaatgtacagctagtgtctgataccattgaggacatagatttggtctgaaagtttgttgtgactaagcagatccgttcttcacgctgaggttcgcgaaacttgagtttctgaggaaacggtggaaagtctttcggcattcgaatgataatcatgtctcttccatctatcagatgttgctgaagcgtcgttgtgtttttgaccttaaagactccatgaagggattgtaccaacagtgtaccattattgcgcctaaagaggtgtttgttcgtgatgatgaatggcccaaagccaatgccatacaagctagtggtatgaccatcactctcattggtcagatggcaaatagtcgagctaatcggattatagtcacgtggtcctttaaacaacgatcttcccattactagagttcacctcctttctctagtatgtgaattgttatccgctcacaattccacacacacattatgccacaccttgtagataaagtcaacaacttttgcaactttctcggcaaaatgtggaattgtgagcgctcacaattccacaaccctcgag
encoding: naseq.html

https://synbiohub.org/public/igem/BBa_B0011_sequence/1
type: Sequence
persistent identity: BBa B0011 sequence
display id: BBa_B0011_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: agagaatataaaaagccagattattaatccggcttttttattattt
encoding: naseq.html

https://synbiohub.org/public/igem/BBa_B0012_sequence/1
type: Sequence
persistent identity: BBa B0012 sequence
display id: BBa_B0012_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: tcacactggctcaccttcgggtgggcctttctgcgtttata
encoding: naseq.html

https://synbiohub.org/public/igem/BBa_K143012_sequence/1
type: Sequence
persistent identity: BBa K143012 sequence
display id: BBa_K143012_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: aattttgtcaaaataattttattgacaacgtcttattaacgttgatataatttaaattttatttgacaaaaatgggctcgtgttgtacaataaatgt
encoding: naseq.html

https://synbiohub.org/public/igem/BBa_K143021_sequence/1
type: Sequence
persistent identity: BBa K143021 sequence
display id: BBa_K143021_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: aaaggtggtgaa
encoding: naseq.html

https://synbiohub.org/public/igem/BBa_K316000_sequence/1
type: Sequence
persistent identity: BBa K316000 sequence
display id: BBa_K316000_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: ttcacctcctttctctagtatgtgaattgttatccgctcacaattccacacacacattatgccacaccttgtagataaagtcaacaacttttgcaactttctcggcaaaatgtggaattgtgagcgctcacaattccacaaccctcgag
encoding: naseq.html

https://synbiohub.org/public/igem/BBa_K316012_sequence/1
type: Sequence
persistent identity: BBa K316012 sequence
display id: BBa_K316012_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: ttattagttcattaactgtgtggcctctttaaccggttgaaacggttcttccggtttatccatgaacactttgtgacctccccaaagaacggaatctgcattcagtctccagcctgaaacccattgttgagcttcttggttcgtcagtaactccatgaagttcttgggaacactcgtaaagtaattattcgtattggtgaagtttgacgcagagtgtatgcccacaataaacccatcccgtgttgacacaagaggagagccacattgcccatcttttgtctgaatccaatgcttccaaaagatgccatcagagctagggaatgtacagctagtgtctgataccattgaggacatagatttggtctgaaagtttgttgtgactaagcagatccgttcttcacgctgaggttcgcgaaacttgagtttctgaggaaacggtggaaagtctttcggcattcgaatgataatcatgtctcttccatctatcagatgttgctgaagcgtcgttgtgtttttgaccttaaagactccatgaagggattgtaccaacagtgtaccattattgcgcctaaagaggtgtttgttcgtgatgatgaatggcccaaagccaatgccatacaagctagtggtatgaccatcactctcattggtcagatggcaaatagtcgagctaatcggattatagtcacgtggtcctttaaacaacgatcttcccat
encoding: naseq.html

iGEM to SBOL conversion
type: Activity
persistent identity: igem2sbol
display id: igem2sbol
version: 1
description: Conversion of the iGEM parts registry to SBOL2.1
owned by: james
owned by: myers
creator: James Alastair McLaughlin
creator: Chris J. Myers
ended at time: 2017-03-06T15:00:00.000Z