//function/crispr/cas9
type: Collection
persistent identity: function crispr cas9
display id: function_crispr_cas9
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
owned by: james
owned by: myers
member: BBa_K1026001
member: BBa_K1994016
member: Prokaryoti
member: BBa_K1150050
member: BBa_K1160000
member: BBa_K1559005
member: BBa_K1137014
member: BBa_K1150017
member: BBa_K1994017
member: BBa_K1150000
member: BBa_K1982007
member: BBa_K1994015
member: BBa_K1946002
member: TC
member: BBa_K1129006
member: BBa_K1179002
member: BBa_K1026000
member: BBa_K1982004
member: BBa_K1559004
member: BBa_K1982008
member: BBa_K1994044
member: BBa_K1559008
member: VP64
member: BBa_K1774001
member: BBa_K1994013
member: BBa_K1559003
member: BBa_K1081000
member: BBa_K1559002
member: BBa_K1559007
member: BBa_K1982011
member: BBa_K1994021
member: BBa_K1218011
member: BBa_K1994025
member: BBa_K1559006
member: BBa_K1982006

Prokaryoti
type: ComponentDefinition
persistent identity: BBa K1982001
display id: BBa_K1982001
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Prokaryotic tCAS9
creator: Zexu Li
created: 2016-09-09T11:00:00Z
modified: 2016-10-09T12:35:07Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2449_
m user id: 30176
owner id: 30176
owning group id: 9
status: status/Planning
bookmark: false
mutable description: tCas9 is a standardized (RFC 10) protein.Interacting with a DNA-binding RNA and fused with different effector domains it can be used for specific gene regulation. Cas9 is the main protein of the CRISPR/Cas system II of Streptococcus pyogenes. CRISPR systems protect bacteria and archaea from phages by recognizing and cleaving of invading phage DNA.This recognition is based on Watson Crick base pairing between a short RNA, called crRNA, and the complementary DNA strand. A second RNA, called tracrRNA, connects crRNA and Cas9. These three parts together form a protein-RNA-DNA complex with the targeted DNA strand [1] Cas9 became of great interest for research concerning DNA targeting, because of its ability to recognize site specific DNA strands by a crRNA. At first the functionality of Cas9 was modified by exchanging aminoacids. As a result, Cas9 was able to introduce mutations within the genome of several organisms by causing double strand breaks [2][3]. Then, it was converted from a nuclease to a nickase introducing single strand breaks [4] and lately it was converted to an enzymatically inactive form, called dCas9 [5]. This tCas9 is standardized (RFC 10). It can be used as a DNA binding protein, that can be fused with different effectors in order to regulate gene expression.
mutable notes: mutant Pst 1 :gctacctgcaG to gctacctgcaC mutant Pst 1 :actatctgcaG to actatctgcaC
mutable provenance: GE Share [1] Westra E.R., Swarts D.C., Staals R.H., Jore M.M., Brouns S.J., van der Oost J. (2012). The CRISPRs, they are a-changin': how prokaryotes generate adaptive immunity. Annu Rev Genet. 46, 311-39 [2] Mali P., Yang L., Esvelt K.M., Aach J., Guell M., DiCarlo J.E., Norville J.E., Church G.M. (2013). RNA-guided human genome engineering via Cas9. Science 339(6121), 823-6 [3] Jiang W., Bikard D., Cox D., Zhang F., Marraffini L.A. (2013). RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat Biotechnol. 31(3), 233-9 [4] Cong, L., Ran, F.A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P.D., Wu, X., Jiang, W., Marraffini, L.A., Zhang, F. (2013). Multiplex Genome Engineering Using CRISPR/Cas Systems. Science 339 (6121), 819-23 [5] Qi L.S., Larson M.H., Gilbert L.A., Doudna J.A., Weissman J.S., Arkin A.P., Lim W.A. (2013). Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. Cell 152(5), 1173-83
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: ATG
sequence annotation: Prokaryotic rbs
sequence: https://synbiohub.org/public/igem/BBa_K1982001_sequence/1

ATG
type: SequenceAnnotation
persistent identity: annotation2482946
display id: annotation2482946
version: 1
top level: Prokaryoti
role: SO:0000318
role: feature/start
location: https://synbiohub.org/public/igem/BBa_K1982001/annotation2482946/range2482946/1

https://synbiohub.org/public/igem/BBa_K1982001/annotation2482946/range2482946/1
type: Range
persistent identity: range2482946
display id: range2482946
version: 1
direction: direction/forward
top level: Prokaryoti
start: 18
end: 21
orientation: inline

Prokaryotic rbs
type: SequenceAnnotation
persistent identity: annotation2488562
display id: annotation2488562
version: 1
top level: Prokaryoti
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K1982001/annotation2488562/range2488562/1

https://synbiohub.org/public/igem/BBa_K1982001/annotation2488562/range2488562/1
type: Range
persistent identity: range2488562
display id: range2488562
version: 1
direction: direction/forward
top level: Prokaryoti
start: 1
end: 18
orientation: inline

BBa_K1129006
type: ComponentDefinition
persistent identity: BBa K1129006
display id: BBa_K1129006
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Cas 9 from Streptococcus thermophilus
creator: UBC iGEM 2013
created: 2013-08-29T11:00:00Z
modified: 2015-05-08T01:09:19Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _1441_
m user id: 0
owner id: 9774
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: true
mutable description: Cas 9 from Streptococcus thermophilus
mutable notes: Illegal cut sites removed
mutable provenance: Streptococcus thermophilus
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: Cas9
sequence: https://synbiohub.org/public/igem/BBa_K1129006_sequence/1

Cas9
type: SequenceAnnotation
persistent identity: annotation2430437
display id: annotation2430437
version: 1
top level: BBa_K1129006
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1129006/annotation2430437/range2430437/1

https://synbiohub.org/public/igem/BBa_K1129006/annotation2430437/range2430437/1
type: Range
persistent identity: range2430437
display id: range2430437
version: 1
direction: direction/forward
top level: BBa_K1129006
start: 1
end: 4167
orientation: inline

BBa_K1994013
type: ComponentDefinition
persistent identity: BBa K1994013
display id: BBa_K1994013
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: sgRNA with dCas9 binding site sequence 9 and PP7 handle insert
creator: Liam Carroll
created: 2016-09-21T11:00:00Z
modified: 2016-10-19T09:23:58Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2461_
m user id: 30870
owner id: 27126
owning group id: 9
status: status/Planning
bookmark: true
mutable description: For use in CRISPR/Cas9 systems. Contains golden gate adapter at 5' end for inserting target sequence. Also contains second golden gate adapter between truncated terminator and aspA terminator. Both golden gate adapters contain Bsa1 cut sites.
mutable notes: No special considerations
mutable provenance: Completely synthetic
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000834
role: partType/RNA
sequence annotation: truncated S. pyrogenes terminator
sequence annotation: 10bp Linker
sequence annotation: dCas9 handle
sequence annotation: aspA terminator
sequence annotation: PP7 BP handle
sequence annotation: Sequence 9
sequence: https://synbiohub.org/public/igem/BBa_K1994013_sequence/1

truncated S. pyrogenes terminator
type: SequenceAnnotation
persistent identity: annotation2484495
display id: annotation2484495
version: 1
top level: BBa_K1994013
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994013/annotation2484495/range2484495/1

https://synbiohub.org/public/igem/BBa_K1994013/annotation2484495/range2484495/1
type: Range
persistent identity: range2484495
display id: range2484495
version: 1
direction: direction/forward
top level: BBa_K1994013
start: 63
end: 96
orientation: inline

10bp Linker
type: SequenceAnnotation
persistent identity: annotation2484496
display id: annotation2484496
version: 1
top level: BBa_K1994013
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994013/annotation2484496/range2484496/1

https://synbiohub.org/public/igem/BBa_K1994013/annotation2484496/range2484496/1
type: Range
persistent identity: range2484496
display id: range2484496
version: 1
direction: direction/forward
top level: BBa_K1994013
start: 97
end: 107
orientation: inline

dCas9 handle
type: SequenceAnnotation
persistent identity: annotation2484494
display id: annotation2484494
version: 1
top level: BBa_K1994013
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994013/annotation2484494/range2484494/1

https://synbiohub.org/public/igem/BBa_K1994013/annotation2484494/range2484494/1
type: Range
persistent identity: range2484494
display id: range2484494
version: 1
direction: direction/forward
top level: BBa_K1994013
start: 21
end: 62
orientation: inline

10bp Linker
type: SequenceAnnotation
persistent identity: annotation2524245
display id: annotation2524245
version: 1
top level: BBa_K1994013
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994013/annotation2524245/range2524245/1

https://synbiohub.org/public/igem/BBa_K1994013/annotation2524245/range2524245/1
type: Range
persistent identity: range2524245
display id: range2524245
version: 1
direction: direction/forward
top level: BBa_K1994013
start: 139
end: 149
orientation: inline

aspA terminator
type: SequenceAnnotation
persistent identity: annotation2484497
display id: annotation2484497
version: 1
top level: BBa_K1994013
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994013/annotation2484497/range2484497/1

https://synbiohub.org/public/igem/BBa_K1994013/annotation2484497/range2484497/1
type: Range
persistent identity: range2484497
display id: range2484497
version: 1
direction: direction/forward
top level: BBa_K1994013
start: 150
end: 189
orientation: inline

PP7 BP handle
type: SequenceAnnotation
persistent identity: annotation2524244
display id: annotation2524244
version: 1
top level: BBa_K1994013
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994013/annotation2524244/range2524244/1

https://synbiohub.org/public/igem/BBa_K1994013/annotation2524244/range2524244/1
type: Range
persistent identity: range2524244
display id: range2524244
version: 1
direction: direction/forward
top level: BBa_K1994013
start: 108
end: 138
orientation: inline

Sequence 9
type: SequenceAnnotation
persistent identity: annotation2484493
display id: annotation2484493
version: 1
top level: BBa_K1994013
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994013/annotation2484493/range2484493/1

https://synbiohub.org/public/igem/BBa_K1994013/annotation2484493/range2484493/1
type: Range
persistent identity: range2484493
display id: range2484493
version: 1
direction: direction/forward
top level: BBa_K1994013
start: 1
end: 20
orientation: inline

BBa_K1559005
type: ComponentDefinition
persistent identity: BBa K1559005
display id: BBa_K1559005
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: CRISPR/Cas9 system with Anderson low-expression constitutive promoter
creator: Xiuqi (Rex) Xia
created: 2014-10-12T11:00:00Z
modified: 2015-05-08T01:10:53Z
discontinued: false
dominant: false
group u list: _1944_
m user id: 0
owner id: 23047
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: TODO
mutable notes: TODO
mutable provenance: TODO
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Generator
component: BBa_J23114
component: BBa_K1559002
component: BBa_B0034
sequence annotation: BBa_J23114
sequence annotation: BBa_B0034
sequence annotation: BBa_K1559002
sequence: https://synbiohub.org/public/igem/BBa_K1559005_sequence/1

BBa_J23114
type: Component
persistent identity: component2418646
display id: component2418646
version: 1
top level: BBa_K1559005
access: public
definition: https://synbiohub.org/public/igem/BBa_J23114/1

BBa_K1559002
type: Component
persistent identity: component2418649
display id: component2418649
version: 1
top level: BBa_K1559005
access: public
definition: BBa_K1559002

BBa_B0034
type: Component
persistent identity: component2418648
display id: component2418648
version: 1
top level: BBa_K1559005
access: public
definition: https://synbiohub.org/public/igem/BBa_B0034/1

BBa_J23114
type: SequenceAnnotation
persistent identity: annotation2418646
display id: annotation2418646
version: 1
top level: BBa_K1559005
location: https://synbiohub.org/public/igem/BBa_K1559005/annotation2418646/range2418646/1
component: BBa_J23114

https://synbiohub.org/public/igem/BBa_K1559005/annotation2418646/range2418646/1
type: Range
persistent identity: range2418646
display id: range2418646
version: 1
direction: direction/reverse
top level: BBa_K1559005
start: 1
end: 35
orientation: inline

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2418648
display id: annotation2418648
version: 1
top level: BBa_K1559005
location: https://synbiohub.org/public/igem/BBa_K1559005/annotation2418648/range2418648/1
component: BBa_B0034

https://synbiohub.org/public/igem/BBa_K1559005/annotation2418648/range2418648/1
type: Range
persistent identity: range2418648
display id: range2418648
version: 1
direction: direction/reverse
top level: BBa_K1559005
start: 36
end: 47
orientation: inline

BBa_K1559002
type: SequenceAnnotation
persistent identity: annotation2418649
display id: annotation2418649
version: 1
top level: BBa_K1559005
location: https://synbiohub.org/public/igem/BBa_K1559005/annotation2418649/range2418649/1
component: BBa_K1559002

https://synbiohub.org/public/igem/BBa_K1559005/annotation2418649/range2418649/1
type: Range
persistent identity: range2418649
display id: range2418649
version: 1
direction: direction/reverse
top level: BBa_K1559005
start: 48
end: 5127
orientation: inline

BBa_K1150000
type: ComponentDefinition
persistent identity: BBa K1150000
display id: BBa_K1150000
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: dCas9
creator: Freiburg 2013
created: 2013-09-14T11:00:00Z
modified: 2015-05-08T01:09:27Z
discontinued: false
dominant: false
experience: experience/None
group u list: _1462_
m user id: 0
owner id: 16627
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable description: The human codon optimized Cas9 from the organism Streptococcus pyogenes was standardized (RFC 25). Additionally we mutated the Nickase function. In the end we engineered a DNA binding protein, that can be fused with different effectors in order to regulate gene expression.
mutable notes: Restriction sites were deleted by primer overhangs. Parts were fused together by fusion PCR. The nickase function was mutated in order to abbolish the demolition of DNA.
mutable provenance: Streptococcus pyogenes, Zhang lab
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: dCas9
sequence annotation: Nickaseless mutation
sequence: https://synbiohub.org/public/igem/BBa_K1150000_sequence/1

dCas9
type: SequenceAnnotation
persistent identity: annotation2347618
display id: annotation2347618
version: 1
top level: BBa_K1150000
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1150000/annotation2347618/range2347618/1

https://synbiohub.org/public/igem/BBa_K1150000/annotation2347618/range2347618/1
type: Range
persistent identity: range2347618
display id: range2347618
version: 1
direction: direction/forward
top level: BBa_K1150000
start: 1
end: 4101
orientation: inline

Nickaseless mutation
type: SequenceAnnotation
persistent identity: annotation2354601
display id: annotation2354601
version: 1
top level: BBa_K1150000
role: SO:0001059
role: feature/mutation
location: https://synbiohub.org/public/igem/BBa_K1150000/annotation2354601/range2354601/1

https://synbiohub.org/public/igem/BBa_K1150000/annotation2354601/range2354601/1
type: Range
persistent identity: range2354601
display id: range2354601
version: 1
direction: direction/forward
top level: BBa_K1150000
start: 2515
end: 2517
orientation: inline

BBa_K1026001
type: ComponentDefinition
persistent identity: BBa K1026001
display id: BBa_K1026001
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: dCas9
creator: Hongyi WU
created: 2013-09-15T11:00:00Z
modified: 2015-05-08T01:08:46Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _1333_
m user id: 0
owner id: 12098
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: dCas9 is a mutant of Cas9, with its endonuclease activity wiped out. dCas9 acts as the sole protein in CRISPRi system (Qi et al., 2013). Reference QI, LEI S., LARSON, MATTHEW H., GILBERT, LUKE A., DOUDNA, JENNIFER A., WEISSMAN, JONATHAN S., ARKIN, ADAM P. & LIM, WENDELL A. 2013. Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell, 152, 1173-1183.
mutable notes: Mutation: Aspartate 10 to Alanine (D10A) and Histidine 840 to Alanine (H840A) in order to cripple the endonuclease activity.
mutable provenance: The dCas9 comes from S. pyogenes.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence: https://synbiohub.org/public/igem/BBa_K1026001_sequence/1

BBa_K1774001
type: ComponentDefinition
persistent identity: BBa K1774001
display id: BBa_K1774001
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Cas9 (optimized for expression in E. coli)
creator: HKU iGEM 2015
created: 2015-09-17T11:00:00Z
modified: 2015-09-20T02:01:59Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2196_
m user id: 25503
owner id: 25503
owning group id: 9
status: status/Planning
bookmark: false
mutable description: This part code for cas9 originated from S. pyogenes SF 370 with codon optimized for expression in E. coli.
mutable notes: Presence of illegal RE sites
mutable provenance: S. pyogenes SF 370
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence: https://synbiohub.org/public/igem/BBa_K1774001_sequence/1

BBa_K1994017
type: ComponentDefinition
persistent identity: BBa K1994017
display id: BBa_K1994017
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: sgRNA with 5' golden gate adapter and PP7 protein binding site
creator: Liam Carroll
created: 2016-09-25T11:00:00Z
modified: 2016-10-19T01:34:26Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2461_
m user id: 30870
owner id: 27126
owning group id: 9
status: status/Planning
bookmark: true
mutable description: For use in CRISPR/Cas9 systems. Contains golden gate adapter at 5' end for inserting target sequence. Contains a PP7 coat protein binding site in the middle.
mutable notes: No special considerations
mutable provenance: Synthetic
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000834
role: partType/RNA
sequence annotation: dCas9 handle
sequence annotation: aspA terminator
sequence annotation: 10bp linker
sequence annotation: truncated S. pyrogenes terminator
sequence annotation: PP7 handle
sequence annotation: 20nt golden gate adapter
sequence: https://synbiohub.org/public/igem/BBa_K1994017_sequence/1

dCas9 handle
type: SequenceAnnotation
persistent identity: annotation2484943
display id: annotation2484943
version: 1
top level: BBa_K1994017
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994017/annotation2484943/range2484943/1

https://synbiohub.org/public/igem/BBa_K1994017/annotation2484943/range2484943/1
type: Range
persistent identity: range2484943
display id: range2484943
version: 1
direction: direction/forward
top level: BBa_K1994017
start: 21
end: 62
orientation: inline

aspA terminator
type: SequenceAnnotation
persistent identity: annotation2484948
display id: annotation2484948
version: 1
top level: BBa_K1994017
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994017/annotation2484948/range2484948/1

https://synbiohub.org/public/igem/BBa_K1994017/annotation2484948/range2484948/1
type: Range
persistent identity: range2484948
display id: range2484948
version: 1
direction: direction/forward
top level: BBa_K1994017
start: 150
end: 178
orientation: inline

10bp linker
type: SequenceAnnotation
persistent identity: annotation2484947
display id: annotation2484947
version: 1
top level: BBa_K1994017
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994017/annotation2484947/range2484947/1

https://synbiohub.org/public/igem/BBa_K1994017/annotation2484947/range2484947/1
type: Range
persistent identity: range2484947
display id: range2484947
version: 1
direction: direction/forward
top level: BBa_K1994017
start: 140
end: 150
orientation: inline

10bp linker
type: SequenceAnnotation
persistent identity: annotation2484945
display id: annotation2484945
version: 1
top level: BBa_K1994017
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994017/annotation2484945/range2484945/1

https://synbiohub.org/public/igem/BBa_K1994017/annotation2484945/range2484945/1
type: Range
persistent identity: range2484945
display id: range2484945
version: 1
direction: direction/forward
top level: BBa_K1994017
start: 97
end: 107
orientation: inline

truncated S. pyrogenes terminator
type: SequenceAnnotation
persistent identity: annotation2484944
display id: annotation2484944
version: 1
top level: BBa_K1994017
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994017/annotation2484944/range2484944/1

https://synbiohub.org/public/igem/BBa_K1994017/annotation2484944/range2484944/1
type: Range
persistent identity: range2484944
display id: range2484944
version: 1
direction: direction/forward
top level: BBa_K1994017
start: 63
end: 96
orientation: inline

PP7 handle
type: SequenceAnnotation
persistent identity: annotation2484946
display id: annotation2484946
version: 1
top level: BBa_K1994017
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994017/annotation2484946/range2484946/1

https://synbiohub.org/public/igem/BBa_K1994017/annotation2484946/range2484946/1
type: Range
persistent identity: range2484946
display id: range2484946
version: 1
direction: direction/forward
top level: BBa_K1994017
start: 108
end: 139
orientation: inline

20nt golden gate adapter
type: SequenceAnnotation
persistent identity: annotation2484942
display id: annotation2484942
version: 1
top level: BBa_K1994017
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994017/annotation2484942/range2484942/1

https://synbiohub.org/public/igem/BBa_K1994017/annotation2484942/range2484942/1
type: Range
persistent identity: range2484942
display id: range2484942
version: 1
direction: direction/forward
top level: BBa_K1994017
start: 1
end: 20
orientation: inline

BBa_K1994015
type: ComponentDefinition
persistent identity: BBa K1994015
display id: BBa_K1994015
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: sgRNA with 5' golden gate adapter and COM protein binding site
creator: Liam Carroll
created: 2016-09-25T11:00:00Z
modified: 2016-10-19T01:28:04Z
discontinued: false
dominant: false
group u list: _2461_
m user id: 30870
owner id: 27126
owning group id: 9
status: status/Planning
bookmark: false
mutable description: For use in CRISPR/Cas9 systems. Contains golden gate adapter at 5' end for inserting target sequence. Contains a COM protein binding site in the middle.
mutable notes: No special considerations
mutable provenance: synthetic
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000834
role: partType/RNA
sequence annotation: COM handle
sequence annotation: truncated S. pyrogenes terminator
sequence annotation: aspA terminator
sequence annotation: 20nt golden gate adapter
sequence annotation: 10bp linker
sequence annotation: dCas9 handle
sequence: https://synbiohub.org/public/igem/BBa_K1994015_sequence/1

COM handle
type: SequenceAnnotation
persistent identity: annotation2484932
display id: annotation2484932
version: 1
top level: BBa_K1994015
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994015/annotation2484932/range2484932/1

https://synbiohub.org/public/igem/BBa_K1994015/annotation2484932/range2484932/1
type: Range
persistent identity: range2484932
display id: range2484932
version: 1
direction: direction/forward
top level: BBa_K1994015
start: 107
end: 125
orientation: inline

truncated S. pyrogenes terminator
type: SequenceAnnotation
persistent identity: annotation2484930
display id: annotation2484930
version: 1
top level: BBa_K1994015
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994015/annotation2484930/range2484930/1

https://synbiohub.org/public/igem/BBa_K1994015/annotation2484930/range2484930/1
type: Range
persistent identity: range2484930
display id: range2484930
version: 1
direction: direction/forward
top level: BBa_K1994015
start: 63
end: 96
orientation: inline

aspA terminator
type: SequenceAnnotation
persistent identity: annotation2484934
display id: annotation2484934
version: 1
top level: BBa_K1994015
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994015/annotation2484934/range2484934/1

https://synbiohub.org/public/igem/BBa_K1994015/annotation2484934/range2484934/1
type: Range
persistent identity: range2484934
display id: range2484934
version: 1
direction: direction/forward
top level: BBa_K1994015
start: 136
end: 176
orientation: inline

20nt golden gate adapter
type: SequenceAnnotation
persistent identity: annotation2484928
display id: annotation2484928
version: 1
top level: BBa_K1994015
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994015/annotation2484928/range2484928/1

https://synbiohub.org/public/igem/BBa_K1994015/annotation2484928/range2484928/1
type: Range
persistent identity: range2484928
display id: range2484928
version: 1
direction: direction/forward
top level: BBa_K1994015
start: 1
end: 20
orientation: inline

10bp linker
type: SequenceAnnotation
persistent identity: annotation2484933
display id: annotation2484933
version: 1
top level: BBa_K1994015
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994015/annotation2484933/range2484933/1

https://synbiohub.org/public/igem/BBa_K1994015/annotation2484933/range2484933/1
type: Range
persistent identity: range2484933
display id: range2484933
version: 1
direction: direction/forward
top level: BBa_K1994015
start: 126
end: 135
orientation: inline

10bp linker
type: SequenceAnnotation
persistent identity: annotation2484931
display id: annotation2484931
version: 1
top level: BBa_K1994015
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994015/annotation2484931/range2484931/1

https://synbiohub.org/public/igem/BBa_K1994015/annotation2484931/range2484931/1
type: Range
persistent identity: range2484931
display id: range2484931
version: 1
direction: direction/forward
top level: BBa_K1994015
start: 97
end: 106
orientation: inline

dCas9 handle
type: SequenceAnnotation
persistent identity: annotation2484929
display id: annotation2484929
version: 1
top level: BBa_K1994015
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994015/annotation2484929/range2484929/1

https://synbiohub.org/public/igem/BBa_K1994015/annotation2484929/range2484929/1
type: Range
persistent identity: range2484929
display id: range2484929
version: 1
direction: direction/forward
top level: BBa_K1994015
start: 21
end: 62
orientation: inline

BBa_K1081000
type: ComponentDefinition
persistent identity: BBa K1081000
display id: BBa_K1081000
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: J13002-dcas9
creator: Hangxing Jia
created: 2013-09-09T11:00:00Z
modified: 2015-05-08T01:09:04Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _1390_
m user id: 0
owner id: 16101
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: true
mutable description: Cas9 is an RNA-guided double-stranded DNA endonuclease, which belongs to the type II bacterial CRISPR system, from Streptococcus pyogenes. Dead cas9(dcas9), in which two mutations were introduced to abolish the endonuclease function, is added behind the biobrick J13002. Here we combine dcas9 with several different guide RNAs to activate or repress one of the double promoters??? transcription.
mutable notes: We made point mutation of EcoRI Restriction Enzyme cutting site inside the coding region of dcas9.
mutable provenance: Dcas9 was provided by addgene. Cas9 is a gene from Streptococcus pyogenes.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Composite
sequence annotation: BBa_J13002
sequence annotation: BBa_K1081000
sequence annotation: R0040
sequence annotation: dcas9
sequence annotation: B0034
sequence: https://synbiohub.org/public/igem/BBa_K1081000_sequence/1

BBa_J13002
type: SequenceAnnotation
persistent identity: annotation2341839
display id: annotation2341839
version: 1
top level: BBa_K1081000
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K1081000/annotation2341839/range2341839/1

https://synbiohub.org/public/igem/BBa_K1081000/annotation2341839/range2341839/1
type: Range
persistent identity: range2341839
display id: range2341839
version: 1
direction: direction/forward
top level: BBa_K1081000
start: 1
end: 74
orientation: inline

BBa_K1081000
type: SequenceAnnotation
persistent identity: annotation2341841
display id: annotation2341841
version: 1
top level: BBa_K1081000
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K1081000/annotation2341841/range2341841/1

https://synbiohub.org/public/igem/BBa_K1081000/annotation2341841/range2341841/1
type: Range
persistent identity: range2341841
display id: range2341841
version: 1
direction: direction/reverse
top level: BBa_K1081000
start: 1
end: 4180
orientation: inline

R0040
type: SequenceAnnotation
persistent identity: annotation2341840
display id: annotation2341840
version: 1
top level: BBa_K1081000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1081000/annotation2341840/range2341840/1

https://synbiohub.org/public/igem/BBa_K1081000/annotation2341840/range2341840/1
type: Range
persistent identity: range2341840
display id: range2341840
version: 1
direction: direction/forward
top level: BBa_K1081000
start: 1
end: 54
orientation: inline

dcas9
type: SequenceAnnotation
persistent identity: annotation2341843
display id: annotation2341843
version: 1
top level: BBa_K1081000
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1081000/annotation2341843/range2341843/1

https://synbiohub.org/public/igem/BBa_K1081000/annotation2341843/range2341843/1
type: Range
persistent identity: range2341843
display id: range2341843
version: 1
direction: direction/forward
top level: BBa_K1081000
start: 77
end: 4180
orientation: inline

B0034
type: SequenceAnnotation
persistent identity: annotation2341842
display id: annotation2341842
version: 1
top level: BBa_K1081000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1081000/annotation2341842/range2341842/1

https://synbiohub.org/public/igem/BBa_K1081000/annotation2341842/range2341842/1
type: Range
persistent identity: range2341842
display id: range2341842
version: 1
direction: direction/forward
top level: BBa_K1081000
start: 63
end: 74
orientation: inline

BBa_K1559002
type: ComponentDefinition
persistent identity: BBa K1559002
display id: BBa_K1559002
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: rearranged CRISPR/Cas9 system without promoter
creator: Xiuqi (Rex) Xia
created: 2014-10-09T11:00:00Z
modified: 2015-05-08T01:10:53Z
discontinued: false
dominant: false
experience: experience/None
group u list: _1944_
m user id: 0
owner id: 23047
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: TODO
mutable notes: TODO
mutable provenance: TODO
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: crRNA array
sequence annotation: spacer placeholder containing BsaI sites
sequence annotation: direct repeat
sequence annotation: predicted terminator rev
sequence annotation: predicted terminator fwd
sequence annotation: tracrRNA
sequence annotation: Region containing bidirectional constitutive promoter
sequence annotation: stem loop within tracrRNA
sequence annotation: cas9
sequence: https://synbiohub.org/public/igem/BBa_K1559002_sequence/1

crRNA array
type: SequenceAnnotation
persistent identity: annotation2418815
display id: annotation2418815
version: 1
top level: BBa_K1559002
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1559002/annotation2418815/range2418815/1

https://synbiohub.org/public/igem/BBa_K1559002/annotation2418815/range2418815/1
type: Range
persistent identity: range2418815
display id: range2418815
version: 1
direction: direction/forward
top level: BBa_K1559002
start: 4260
end: 4361
orientation: inline

spacer placeholder containing BsaI sites
type: SequenceAnnotation
persistent identity: annotation2418813
display id: annotation2418813
version: 1
top level: BBa_K1559002
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1559002/annotation2418813/range2418813/1

https://synbiohub.org/public/igem/BBa_K1559002/annotation2418813/range2418813/1
type: Range
persistent identity: range2418813
display id: range2418813
version: 1
direction: direction/forward
top level: BBa_K1559002
start: 4296
end: 4325
orientation: inline

direct repeat
type: SequenceAnnotation
persistent identity: annotation2418812
display id: annotation2418812
version: 1
top level: BBa_K1559002
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1559002/annotation2418812/range2418812/1

https://synbiohub.org/public/igem/BBa_K1559002/annotation2418812/range2418812/1
type: Range
persistent identity: range2418812
display id: range2418812
version: 1
direction: direction/reverse
top level: BBa_K1559002
start: 4260
end: 4295
orientation: inline

direct repeat
type: SequenceAnnotation
persistent identity: annotation2418814
display id: annotation2418814
version: 1
top level: BBa_K1559002
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1559002/annotation2418814/range2418814/1

https://synbiohub.org/public/igem/BBa_K1559002/annotation2418814/range2418814/1
type: Range
persistent identity: range2418814
display id: range2418814
version: 1
direction: direction/forward
top level: BBa_K1559002
start: 4326
end: 4361
orientation: inline

predicted terminator rev
type: SequenceAnnotation
persistent identity: annotation2425010
display id: annotation2425010
version: 1
top level: BBa_K1559002
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1559002/annotation2425010/range2425010/1

https://synbiohub.org/public/igem/BBa_K1559002/annotation2425010/range2425010/1
type: Range
persistent identity: range2425010
display id: range2425010
version: 1
direction: direction/reverse
top level: BBa_K1559002
start: 4628
end: 4674
orientation: inline

predicted terminator fwd
type: SequenceAnnotation
persistent identity: annotation2425031
display id: annotation2425031
version: 1
top level: BBa_K1559002
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1559002/annotation2425031/range2425031/1

https://synbiohub.org/public/igem/BBa_K1559002/annotation2425031/range2425031/1
type: Range
persistent identity: range2425031
display id: range2425031
version: 1
direction: direction/forward
top level: BBa_K1559002
start: 4628
end: 4676
orientation: inline

tracrRNA
type: SequenceAnnotation
persistent identity: annotation2418816
display id: annotation2418816
version: 1
top level: BBa_K1559002
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1559002/annotation2418816/range2418816/1

https://synbiohub.org/public/igem/BBa_K1559002/annotation2418816/range2418816/1
type: Range
persistent identity: range2418816
display id: range2418816
version: 1
direction: direction/reverse
top level: BBa_K1559002
start: 4700
end: 4870
orientation: inline

Region containing bidirectional constitutive promoter
type: SequenceAnnotation
persistent identity: annotation2418817
display id: annotation2418817
version: 1
top level: BBa_K1559002
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1559002/annotation2418817/range2418817/1

https://synbiohub.org/public/igem/BBa_K1559002/annotation2418817/range2418817/1
type: Range
persistent identity: range2418817
display id: range2418817
version: 1
direction: direction/forward
top level: BBa_K1559002
start: 4871
end: 5080
orientation: inline

stem loop within tracrRNA
type: SequenceAnnotation
persistent identity: annotation2425040
display id: annotation2425040
version: 1
top level: BBa_K1559002
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1559002/annotation2425040/range2425040/1

https://synbiohub.org/public/igem/BBa_K1559002/annotation2425040/range2425040/1
type: Range
persistent identity: range2425040
display id: range2425040
version: 1
direction: direction/reverse
top level: BBa_K1559002
start: 4692
end: 4733
orientation: inline

cas9
type: SequenceAnnotation
persistent identity: annotation2418811
display id: annotation2418811
version: 1
top level: BBa_K1559002
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1559002/annotation2418811/range2418811/1

https://synbiohub.org/public/igem/BBa_K1559002/annotation2418811/range2418811/1
type: Range
persistent identity: range2418811
display id: range2418811
version: 1
direction: direction/forward
top level: BBa_K1559002
start: 1
end: 4107
orientation: inline

BBa_K1559006
type: ComponentDefinition
persistent identity: BBa K1559006
display id: BBa_K1559006
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: CRISPR/Cas9 system with pBAD inducible promoter
creator: Xiuqi (Rex) Xia
created: 2014-10-12T11:00:00Z
modified: 2015-05-08T01:10:53Z
discontinued: false
dominant: false
group u list: _1944_
m user id: 0
owner id: 23047
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: TODO
mutable notes: TODO
mutable provenance: TODO
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Generator
component: BBa_I13453
component: BBa_K1559002
component: BBa_B0034
sequence annotation: BBa_B0034
sequence annotation: BBa_I13453
sequence annotation: BBa_K1559002
sequence: https://synbiohub.org/public/igem/BBa_K1559006_sequence/1

BBa_I13453
type: Component
persistent identity: component2418654
display id: component2418654
version: 1
top level: BBa_K1559006
access: public
definition: https://synbiohub.org/public/igem/BBa_I13453/1

BBa_K1559002
type: Component
persistent identity: component2418657
display id: component2418657
version: 1
top level: BBa_K1559006
access: public
definition: BBa_K1559002

BBa_B0034
type: Component
persistent identity: component2418656
display id: component2418656
version: 1
top level: BBa_K1559006
access: public
definition: https://synbiohub.org/public/igem/BBa_B0034/1

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2418656
display id: annotation2418656
version: 1
top level: BBa_K1559006
location: https://synbiohub.org/public/igem/BBa_K1559006/annotation2418656/range2418656/1
component: BBa_B0034

https://synbiohub.org/public/igem/BBa_K1559006/annotation2418656/range2418656/1
type: Range
persistent identity: range2418656
display id: range2418656
version: 1
direction: direction/reverse
top level: BBa_K1559006
start: 131
end: 142
orientation: inline

BBa_I13453
type: SequenceAnnotation
persistent identity: annotation2418654
display id: annotation2418654
version: 1
top level: BBa_K1559006
location: https://synbiohub.org/public/igem/BBa_K1559006/annotation2418654/range2418654/1
component: BBa_I13453

https://synbiohub.org/public/igem/BBa_K1559006/annotation2418654/range2418654/1
type: Range
persistent identity: range2418654
display id: range2418654
version: 1
direction: direction/reverse
top level: BBa_K1559006
start: 1
end: 130
orientation: inline

BBa_K1559002
type: SequenceAnnotation
persistent identity: annotation2418657
display id: annotation2418657
version: 1
top level: BBa_K1559006
location: https://synbiohub.org/public/igem/BBa_K1559006/annotation2418657/range2418657/1
component: BBa_K1559002

https://synbiohub.org/public/igem/BBa_K1559006/annotation2418657/range2418657/1
type: Range
persistent identity: range2418657
display id: range2418657
version: 1
direction: direction/reverse
top level: BBa_K1559006
start: 143
end: 5222
orientation: inline

BBa_K1137014
type: ComponentDefinition
persistent identity: BBa K1137014
display id: BBa_K1137014
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: tracRNA-CAS9
creator: Nicolas Koutsoubelis, Anne L??chner
created: 2013-10-03T11:00:00Z
modified: 2015-05-08T01:09:21Z
discontinued: false
dominant: false
experience: experience/None
group u list: _1449_
m user id: 0
owner id: 11926
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: This complex part contains the tracrRNA and the CAS9 under the control of constitutive promoters. Note that there is no RBS before the tracRNA. The tracrRNA-CAS9 part is the heart of the CRISPR system. The tracrRNA and the corresponding target crRNA dimerize and are processed and hybridize before binding to the CAS9 protein. The tracrRNA has no specific part that is responsible for binding to the targeted DNA. Therefor this part can be used together with any crRNA. The CAS9 gene encodes for the CRISPR associated protein 9 that is needed to generate a double strand break at a target site. CAS9 binds to the tracRNA-crRNA dimer/gRNA and is then guided by them/it to the target site where it generate a double strand break. This part can be used together with the crRNA anti KAN or the gRNA anti KAN and cause double strand breaks. It can of course also be used together with new generated crRNAs/gRNAs. The tracRNA-CAS9 part is taken from the CRISPER plasmid of the Jiang et al. 2013 paper.
mutable notes: -
mutable provenance: The plasmid was ordered from Addgene and come originally from the paper below: Wenyan Jiang, David Bikard, David Cox, Feng Zhang & Luciano A Marraffini (2013): RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nature Biotechnology 31 (3), 233-239.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence: https://synbiohub.org/public/igem/BBa_K1137014_sequence/1

BBa_K1946002
type: ComponentDefinition
persistent identity: BBa K1946002
display id: BBa_K1946002
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: sgRNA targeting LacI
creator: Musa Efe Işılak
created: 2016-10-12T11:00:00Z
modified: 2016-10-14T06:17:48Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _2413_
m user id: 29843
owner id: 29843
owning group id: 9
status: status/Planning
bookmark: false
mutable description: This part has a sgRNA targeting lacI. It also has 5' and 3' ribozyme which cleave the RNA molecule with self catalysis so that any sequence can be added to 5' or 3' to this part without preventing Cas9 binding and its activity.
mutable notes: .
mutable provenance: This part comes from the paper Liu, Yuchen, Yayue Zeng, Li Liu, Chengle Zhuang, Xing Fu, Weiren Huang, and Zhiming Cai. "Synthesizing AND Gate Genetic Circuits Based on CRISPR-Cas9 for Identification of Bladder Cancer Cells." Nature Communications Nat Comms 5 (2014): 5393.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000834
role: partType/RNA
sequence annotation: Cas9 recognition site
sequence annotation: 3' ribozyme
sequence annotation: 5
sequence annotation: LacI targeting region
sequence: https://synbiohub.org/public/igem/BBa_K1946002_sequence/1

Cas9 recognition site
type: SequenceAnnotation
persistent identity: annotation2497327
display id: annotation2497327
version: 1
top level: BBa_K1946002
role: feature/binding
role: SO:0001091
location: https://synbiohub.org/public/igem/BBa_K1946002/annotation2497327/range2497327/1

https://synbiohub.org/public/igem/BBa_K1946002/annotation2497327/range2497327/1
type: Range
persistent identity: range2497327
display id: range2497327
version: 1
direction: direction/forward
top level: BBa_K1946002
start: 79
end: 162
orientation: inline

3' ribozyme
type: SequenceAnnotation
persistent identity: annotation2497329
display id: annotation2497329
version: 1
top level: BBa_K1946002
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1946002/annotation2497329/range2497329/1

https://synbiohub.org/public/igem/BBa_K1946002/annotation2497329/range2497329/1
type: Range
persistent identity: range2497329
display id: range2497329
version: 1
direction: direction/forward
top level: BBa_K1946002
start: 163
end: 205
orientation: inline

5
type: SequenceAnnotation
persistent identity: annotation2497285
display id: annotation2497285
version: 1
top level: BBa_K1946002
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1946002/annotation2497285/range2497285/1

https://synbiohub.org/public/igem/BBa_K1946002/annotation2497285/range2497285/1
type: Range
persistent identity: range2497285
display id: range2497285
version: 1
direction: direction/forward
top level: BBa_K1946002
start: 1
end: 58
orientation: inline

LacI targeting region
type: SequenceAnnotation
persistent identity: annotation2497324
display id: annotation2497324
version: 1
top level: BBa_K1946002
role: feature/binding
role: SO:0001091
location: https://synbiohub.org/public/igem/BBa_K1946002/annotation2497324/range2497324/1

https://synbiohub.org/public/igem/BBa_K1946002/annotation2497324/range2497324/1
type: Range
persistent identity: range2497324
display id: range2497324
version: 1
direction: direction/forward
top level: BBa_K1946002
start: 59
end: 78
orientation: inline

BBa_K1026000
type: ComponentDefinition
persistent identity: BBa K1026000
display id: BBa_K1026000
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Constitutively Expressed dCas9 Operon
creator: Hongyi WU
created: 2013-09-15T11:00:00Z
modified: 2015-05-08T01:08:46Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _1333_
m user id: 0
owner id: 12098
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: true
mutable description: This Constitutively Expressed dCas9 Operon is an assembly of the constitutive promoter BBa_J23100, a strong RBS BBa_B0034, coding sequence of dCas9 protein and a reliable terminator BBa_B0015. dCas9 is a mutant of Cas9, with its endonuclease activity wiped out. dCas9 acts as the sole protein in CRISPRi system (Qi et al., 2013). Reference QI, LEI S., LARSON, MATTHEW H., GILBERT, LUKE A., DOUDNA, JENNIFER A., WEISSMAN, JONATHAN S., ARKIN, ADAM P. & LIM, WENDELL A. 2013. Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell, 152, 1173-1183.
mutable notes: The expression level of dCas9 shall be delicately controlled, so as to acquire continuous adjustment (regulation) of target genes.
mutable provenance: The dCas9 comes from S. pyogenes.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Composite
component: BBa_B0015
component: BBa_B0034
component: BBa_K1026001
component: BBa_J23100
sequence annotation: BBa_B0015
sequence annotation: BBa_K1026001
sequence annotation: BBa_B0034
sequence annotation: BBa_J23100
sequence: https://synbiohub.org/public/igem/BBa_K1026000_sequence/1

BBa_B0015
type: Component
persistent identity: component2347329
display id: component2347329
version: 1
top level: BBa_K1026000
access: public
definition: https://synbiohub.org/public/igem/BBa_B0015/1

BBa_B0034
type: Component
persistent identity: component2347321
display id: component2347321
version: 1
top level: BBa_K1026000
access: public
definition: https://synbiohub.org/public/igem/BBa_B0034/1

BBa_K1026001
type: Component
persistent identity: component2347322
display id: component2347322
version: 1
top level: BBa_K1026000
access: public
definition: BBa_K1026001

BBa_J23100
type: Component
persistent identity: component2347319
display id: component2347319
version: 1
top level: BBa_K1026000
access: public
definition: https://synbiohub.org/public/igem/BBa_J23100/1

BBa_B0015
type: SequenceAnnotation
persistent identity: annotation2347329
display id: annotation2347329
version: 1
top level: BBa_K1026000
location: https://synbiohub.org/public/igem/BBa_K1026000/annotation2347329/range2347329/1
component: BBa_B0015

https://synbiohub.org/public/igem/BBa_K1026000/annotation2347329/range2347329/1
type: Range
persistent identity: range2347329
display id: range2347329
version: 1
direction: direction/reverse
top level: BBa_K1026000
start: 4183
end: 4311
orientation: inline

BBa_K1026001
type: SequenceAnnotation
persistent identity: annotation2347322
display id: annotation2347322
version: 1
top level: BBa_K1026000
location: https://synbiohub.org/public/igem/BBa_K1026000/annotation2347322/range2347322/1
component: BBa_K1026001

https://synbiohub.org/public/igem/BBa_K1026000/annotation2347322/range2347322/1
type: Range
persistent identity: range2347322
display id: range2347322
version: 1
direction: direction/reverse
top level: BBa_K1026000
start: 62
end: 4174
orientation: inline

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2347321
display id: annotation2347321
version: 1
top level: BBa_K1026000
location: https://synbiohub.org/public/igem/BBa_K1026000/annotation2347321/range2347321/1
component: BBa_B0034

https://synbiohub.org/public/igem/BBa_K1026000/annotation2347321/range2347321/1
type: Range
persistent identity: range2347321
display id: range2347321
version: 1
direction: direction/reverse
top level: BBa_K1026000
start: 44
end: 55
orientation: inline

BBa_J23100
type: SequenceAnnotation
persistent identity: annotation2347319
display id: annotation2347319
version: 1
top level: BBa_K1026000
location: https://synbiohub.org/public/igem/BBa_K1026000/annotation2347319/range2347319/1
component: BBa_J23100

https://synbiohub.org/public/igem/BBa_K1026000/annotation2347319/range2347319/1
type: Range
persistent identity: range2347319
display id: range2347319
version: 1
direction: direction/reverse
top level: BBa_K1026000
start: 1
end: 35
orientation: inline

BBa_K1994021
type: ComponentDefinition
persistent identity: BBa K1994021
display id: BBa_K1994021
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: sgRNA containing two golden gate adapters
creator: Isobel Holden
created: 2016-10-13T11:00:00Z
modified: 2016-10-19T08:03:43Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2461_
m user id: 30870
owner id: 30870
owning group id: 9
status: status/Planning
bookmark: true
mutable description: sgRNA with Bsa 1 Golden Gate adaptors
mutable notes: To simplify the traditional gRNA comprised of a crRNA containing a dCas9 handle, and tracrRNA which binds to the PAM sequence in CRISPR, we created a joined sgRNA. As modifying the 3' end of the gRNA does not affect dCas9 binding we have used a deactivated dcas9 terminator and placed golden gate adaptable sites. The two sites are located where the complementary PAM-proximal sequence is located, allowing the distance dCas9 binds from the promoter to be adapted.The other golden gate adaptor in the position after the deactivated terminator so any RNA sequence such as an RNA binding protein handle can be inserted. This region is then followed by a real terminator for the strand.
mutable provenance: Synthetic modification of crRNA and tracrRNA found in S. pyrogenes
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000834
role: partType/RNA
sequence annotation: Golden Gate adaptor
sequence annotation: dCas9 handle
sequence annotation: deactivated dCas9 terminator
sequence annotation: aspA terminator
sequence: https://synbiohub.org/public/igem/BBa_K1994021_sequence/1

Golden Gate adaptor
type: SequenceAnnotation
persistent identity: annotation2503056
display id: annotation2503056
version: 1
top level: BBa_K1994021
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994021/annotation2503056/range2503056/1

https://synbiohub.org/public/igem/BBa_K1994021/annotation2503056/range2503056/1
type: Range
persistent identity: range2503056
display id: range2503056
version: 1
direction: direction/forward
top level: BBa_K1994021
start: 97
end: 116
orientation: inline

dCas9 handle
type: SequenceAnnotation
persistent identity: annotation2503015
display id: annotation2503015
version: 1
top level: BBa_K1994021
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994021/annotation2503015/range2503015/1

https://synbiohub.org/public/igem/BBa_K1994021/annotation2503015/range2503015/1
type: Range
persistent identity: range2503015
display id: range2503015
version: 1
direction: direction/forward
top level: BBa_K1994021
start: 21
end: 62
orientation: inline

Golden Gate adaptor
type: SequenceAnnotation
persistent identity: annotation2502949
display id: annotation2502949
version: 1
top level: BBa_K1994021
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994021/annotation2502949/range2502949/1

https://synbiohub.org/public/igem/BBa_K1994021/annotation2502949/range2502949/1
type: Range
persistent identity: range2502949
display id: range2502949
version: 1
direction: direction/forward
top level: BBa_K1994021
start: 1
end: 20
orientation: inline

deactivated dCas9 terminator
type: SequenceAnnotation
persistent identity: annotation2503036
display id: annotation2503036
version: 1
top level: BBa_K1994021
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994021/annotation2503036/range2503036/1

https://synbiohub.org/public/igem/BBa_K1994021/annotation2503036/range2503036/1
type: Range
persistent identity: range2503036
display id: range2503036
version: 1
direction: direction/forward
top level: BBa_K1994021
start: 63
end: 96
orientation: inline

aspA terminator
type: SequenceAnnotation
persistent identity: annotation2503090
display id: annotation2503090
version: 1
top level: BBa_K1994021
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994021/annotation2503090/range2503090/1

https://synbiohub.org/public/igem/BBa_K1994021/annotation2503090/range2503090/1
type: Range
persistent identity: range2503090
display id: range2503090
version: 1
direction: direction/forward
top level: BBa_K1994021
start: 116
end: 157
orientation: inline

BBa_K1994025
type: ComponentDefinition
persistent identity: BBa K1994025
display id: BBa_K1994025
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: BsaI-GFP-dCas9
creator: Egheosa Ogbomo
created: 2016-10-13T11:00:00Z
modified: 2016-10-19T08:26:08Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2461_
m user id: 30870
owner id: 28134
owning group id: 9
status: status/Planning
bookmark: false
mutable description: GFP-dCas9
mutable notes: GFP-dCas9
mutable provenance: GFP-dCas9
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Composite
component: BBa_K1994044
component: BBa_K1994005
component: BBa_K1994045
component: BBa_K1994004
component: BBa_K1994002
component: BBa_K1994003
component: BBa_K1994000
sequence annotation: BBa_K1994002
sequence annotation: BBa_K1994004
sequence annotation: BBa_K1994044
sequence annotation: BBa_K1994000
sequence annotation: BBa_K1994005
sequence annotation: BBa_K1994045
sequence annotation: BBa_K1994003
sequence: https://synbiohub.org/public/igem/BBa_K1994025_sequence/1

BBa_K1994044
type: Component
persistent identity: component2523959
display id: component2523959
version: 1
top level: BBa_K1994025
access: public
definition: BBa_K1994044

BBa_K1994005
type: Component
persistent identity: component2523948
display id: component2523948
version: 1
top level: BBa_K1994025
access: public
definition: https://synbiohub.org/public/igem/BBa_K1994005/1

BBa_K1994045
type: Component
persistent identity: component2523956
display id: component2523956
version: 1
top level: BBa_K1994025
access: public
definition: https://synbiohub.org/public/igem/BBa_K1994045/1

BBa_K1994004
type: Component
persistent identity: component2523951
display id: component2523951
version: 1
top level: BBa_K1994025
access: public
definition: https://synbiohub.org/public/igem/BBa_K1994004/1

BBa_K1994002
type: Component
persistent identity: component2523960
display id: component2523960
version: 1
top level: BBa_K1994025
access: public
definition: https://synbiohub.org/public/igem/BBa_K1994002/1

BBa_K1994003
type: Component
persistent identity: component2523952
display id: component2523952
version: 1
top level: BBa_K1994025
access: public
definition: https://synbiohub.org/public/igem/BBa_K1994003/1

BBa_K1994000
type: Component
persistent identity: component2523961
display id: component2523961
version: 1
top level: BBa_K1994025
access: public
definition: https://synbiohub.org/public/igem/BBa_K1994000/1

BBa_K1994002
type: SequenceAnnotation
persistent identity: annotation2523960
display id: annotation2523960
version: 1
top level: BBa_K1994025
location: https://synbiohub.org/public/igem/BBa_K1994025/annotation2523960/range2523960/1
component: BBa_K1994002

https://synbiohub.org/public/igem/BBa_K1994025/annotation2523960/range2523960/1
type: Range
persistent identity: range2523960
display id: range2523960
version: 1
direction: direction/reverse
top level: BBa_K1994025
start: 934
end: 963
orientation: inline

BBa_K1994004
type: SequenceAnnotation
persistent identity: annotation2523951
display id: annotation2523951
version: 1
top level: BBa_K1994025
location: https://synbiohub.org/public/igem/BBa_K1994025/annotation2523951/range2523951/1
component: BBa_K1994004

https://synbiohub.org/public/igem/BBa_K1994025/annotation2523951/range2523951/1
type: Range
persistent identity: range2523951
display id: range2523951
version: 1
direction: direction/reverse
top level: BBa_K1994025
start: 29
end: 75
orientation: inline

BBa_K1994044
type: SequenceAnnotation
persistent identity: annotation2523959
display id: annotation2523959
version: 1
top level: BBa_K1994025
location: https://synbiohub.org/public/igem/BBa_K1994025/annotation2523959/range2523959/1
component: BBa_K1994044

https://synbiohub.org/public/igem/BBa_K1994025/annotation2523959/range2523959/1
type: Range
persistent identity: range2523959
display id: range2523959
version: 1
direction: direction/reverse
top level: BBa_K1994025
start: 878
end: 925
orientation: inline

BBa_K1994000
type: SequenceAnnotation
persistent identity: annotation2523961
display id: annotation2523961
version: 1
top level: BBa_K1994025
location: https://synbiohub.org/public/igem/BBa_K1994025/annotation2523961/range2523961/1
component: BBa_K1994000

https://synbiohub.org/public/igem/BBa_K1994025/annotation2523961/range2523961/1
type: Range
persistent identity: range2523961
display id: range2523961
version: 1
direction: direction/reverse
top level: BBa_K1994025
start: 972
end: 5075
orientation: inline

BBa_K1994005
type: SequenceAnnotation
persistent identity: annotation2523948
display id: annotation2523948
version: 1
top level: BBa_K1994025
location: https://synbiohub.org/public/igem/BBa_K1994025/annotation2523948/range2523948/1
component: BBa_K1994005

https://synbiohub.org/public/igem/BBa_K1994025/annotation2523948/range2523948/1
type: Range
persistent identity: range2523948
display id: range2523948
version: 1
direction: direction/reverse
top level: BBa_K1994025
start: 1
end: 20
orientation: inline

BBa_K1994045
type: SequenceAnnotation
persistent identity: annotation2523956
display id: annotation2523956
version: 1
top level: BBa_K1994025
location: https://synbiohub.org/public/igem/BBa_K1994025/annotation2523956/range2523956/1
component: BBa_K1994045

https://synbiohub.org/public/igem/BBa_K1994025/annotation2523956/range2523956/1
type: Range
persistent identity: range2523956
display id: range2523956
version: 1
direction: direction/reverse
top level: BBa_K1994025
start: 813
end: 869
orientation: inline

BBa_K1994003
type: SequenceAnnotation
persistent identity: annotation2523952
display id: annotation2523952
version: 1
top level: BBa_K1994025
location: https://synbiohub.org/public/igem/BBa_K1994025/annotation2523952/range2523952/1
component: BBa_K1994003

https://synbiohub.org/public/igem/BBa_K1994025/annotation2523952/range2523952/1
type: Range
persistent identity: range2523952
display id: range2523952
version: 1
direction: direction/reverse
top level: BBa_K1994025
start: 82
end: 804
orientation: inline

BBa_K1994016
type: ComponentDefinition
persistent identity: BBa K1994016
display id: BBa_K1994016
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: sgRNA with 5' golden gate adapter and MS2 coat protein binding site
creator: Liam Carroll
created: 2016-09-25T11:00:00Z
modified: 2016-10-19T01:26:31Z
discontinued: false
dominant: false
group u list: _2461_
m user id: 30870
owner id: 27126
owning group id: 9
status: status/Planning
bookmark: false
mutable description: For use in CRISPR/Cas9 systems. Contains golden gate adapter at 5' end for inserting target sequence. Contains a MS2 coat protein binding site in the middle.
mutable notes: No special considerations
mutable provenance: Synthetic
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000834
role: partType/RNA
sequence annotation: 5bp linker
sequence annotation: MS2 handle
sequence annotation: dCas9 handle
sequence annotation: 20nt golden gate adapter
sequence annotation: truncated S. pyrogenes terminator
sequence annotation: aspA terminator
sequence: https://synbiohub.org/public/igem/BBa_K1994016_sequence/1

5bp linker
type: SequenceAnnotation
persistent identity: annotation2484940
display id: annotation2484940
version: 1
top level: BBa_K1994016
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994016/annotation2484940/range2484940/1

https://synbiohub.org/public/igem/BBa_K1994016/annotation2484940/range2484940/1
type: Range
persistent identity: range2484940
display id: range2484940
version: 1
direction: direction/forward
top level: BBa_K1994016
start: 127
end: 131
orientation: inline

MS2 handle
type: SequenceAnnotation
persistent identity: annotation2484939
display id: annotation2484939
version: 1
top level: BBa_K1994016
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994016/annotation2484939/range2484939/1

https://synbiohub.org/public/igem/BBa_K1994016/annotation2484939/range2484939/1
type: Range
persistent identity: range2484939
display id: range2484939
version: 1
direction: direction/forward
top level: BBa_K1994016
start: 102
end: 126
orientation: inline

5bp linker
type: SequenceAnnotation
persistent identity: annotation2484938
display id: annotation2484938
version: 1
top level: BBa_K1994016
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994016/annotation2484938/range2484938/1

https://synbiohub.org/public/igem/BBa_K1994016/annotation2484938/range2484938/1
type: Range
persistent identity: range2484938
display id: range2484938
version: 1
direction: direction/forward
top level: BBa_K1994016
start: 97
end: 101
orientation: inline

dCas9 handle
type: SequenceAnnotation
persistent identity: annotation2484936
display id: annotation2484936
version: 1
top level: BBa_K1994016
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994016/annotation2484936/range2484936/1

https://synbiohub.org/public/igem/BBa_K1994016/annotation2484936/range2484936/1
type: Range
persistent identity: range2484936
display id: range2484936
version: 1
direction: direction/forward
top level: BBa_K1994016
start: 21
end: 62
orientation: inline

20nt golden gate adapter
type: SequenceAnnotation
persistent identity: annotation2484935
display id: annotation2484935
version: 1
top level: BBa_K1994016
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994016/annotation2484935/range2484935/1

https://synbiohub.org/public/igem/BBa_K1994016/annotation2484935/range2484935/1
type: Range
persistent identity: range2484935
display id: range2484935
version: 1
direction: direction/forward
top level: BBa_K1994016
start: 1
end: 20
orientation: inline

truncated S. pyrogenes terminator
type: SequenceAnnotation
persistent identity: annotation2484937
display id: annotation2484937
version: 1
top level: BBa_K1994016
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K1994016/annotation2484937/range2484937/1

https://synbiohub.org/public/igem/BBa_K1994016/annotation2484937/range2484937/1
type: Range
persistent identity: range2484937
display id: range2484937
version: 1
direction: direction/forward
top level: BBa_K1994016
start: 63
end: 96
orientation: inline

aspA terminator
type: SequenceAnnotation
persistent identity: annotation2484941
display id: annotation2484941
version: 1
top level: BBa_K1994016
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994016/annotation2484941/range2484941/1

https://synbiohub.org/public/igem/BBa_K1994016/annotation2484941/range2484941/1
type: Range
persistent identity: range2484941
display id: range2484941
version: 1
direction: direction/forward
top level: BBa_K1994016
start: 132
end: 172
orientation: inline

BBa_K1559008
type: ComponentDefinition
persistent identity: BBa K1559008
display id: BBa_K1559008
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: CRISPR/Cas9 system with pRha inducible promoter
creator: Xiuqi (Rex) Xia
created: 2014-10-12T11:00:00Z
modified: 2015-05-08T01:10:53Z
discontinued: false
dominant: false
group u list: _1944_
m user id: 0
owner id: 23047
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: TODO
mutable notes: TODO
mutable provenance: TODO
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Generator
component: BBa_B0034
component: BBa_K1559002
component: BBa_K914003
sequence annotation: BBa_B0034
sequence annotation: BBa_K914003
sequence annotation: BBa_K1559002
sequence: https://synbiohub.org/public/igem/BBa_K1559008_sequence/1

BBa_B0034
type: Component
persistent identity: component2418693
display id: component2418693
version: 1
top level: BBa_K1559008
access: public
definition: https://synbiohub.org/public/igem/BBa_B0034/1

BBa_K1559002
type: Component
persistent identity: component2418694
display id: component2418694
version: 1
top level: BBa_K1559008
access: public
definition: BBa_K1559002

BBa_K914003
type: Component
persistent identity: component2418691
display id: component2418691
version: 1
top level: BBa_K1559008
access: public
definition: https://synbiohub.org/public/igem/BBa_K914003/1

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2418693
display id: annotation2418693
version: 1
top level: BBa_K1559008
location: https://synbiohub.org/public/igem/BBa_K1559008/annotation2418693/range2418693/1
component: BBa_B0034

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type: Range
persistent identity: range2418693
display id: range2418693
version: 1
direction: direction/reverse
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start: 123
end: 134
orientation: inline

BBa_K914003
type: SequenceAnnotation
persistent identity: annotation2418691
display id: annotation2418691
version: 1
top level: BBa_K1559008
location: https://synbiohub.org/public/igem/BBa_K1559008/annotation2418691/range2418691/1
component: BBa_K914003

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type: Range
persistent identity: range2418691
display id: range2418691
version: 1
direction: direction/reverse
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start: 1
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orientation: inline

BBa_K1559002
type: SequenceAnnotation
persistent identity: annotation2418694
display id: annotation2418694
version: 1
top level: BBa_K1559008
location: https://synbiohub.org/public/igem/BBa_K1559008/annotation2418694/range2418694/1
component: BBa_K1559002

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type: Range
persistent identity: range2418694
display id: range2418694
version: 1
direction: direction/reverse
top level: BBa_K1559008
start: 135
end: 5214
orientation: inline

BBa_K1179002
type: ComponentDefinition
persistent identity: BBa K1179002
display id: BBa_K1179002
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Hef1A_Cas9-VP16
creator: Brandon Nadres
created: 2013-09-16T11:00:00Z
modified: 2015-05-08T01:09:35Z
discontinued: false
dominant: false
experience: experience/None
group u list: _1492_
m user id: 0
owner id: 18179
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: true
mutable description: This part is submitted under the RFC 65 standard. Stay tuned for more description, the MIT 2013 Team.
mutable notes: hi
mutable provenance: Source
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Generator
component: BBa_J85600
component: BBa_K1179009
component: BBa_K779200
sequence annotation: BBa_K779200
sequence annotation: BBa_J85600
sequence annotation: BBa_K1179009
sequence: https://synbiohub.org/public/igem/BBa_K1179002_sequence/1

BBa_J85600
type: Component
persistent identity: component2353737
display id: component2353737
version: 1
top level: BBa_K1179002
access: public
definition: https://synbiohub.org/public/igem/BBa_J85600/1

BBa_K1179009
type: Component
persistent identity: component2353738
display id: component2353738
version: 1
top level: BBa_K1179002
access: public
definition: https://synbiohub.org/public/igem/BBa_K1179009/1

BBa_K779200
type: Component
persistent identity: component2353735
display id: component2353735
version: 1
top level: BBa_K1179002
access: public
definition: https://synbiohub.org/public/igem/BBa_K779200/1

BBa_K779200
type: SequenceAnnotation
persistent identity: annotation2353735
display id: annotation2353735
version: 1
top level: BBa_K1179002
location: https://synbiohub.org/public/igem/BBa_K1179002/annotation2353735/range2353735/1
component: BBa_K779200

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type: Range
persistent identity: range2353735
display id: range2353735
version: 1
direction: direction/reverse
top level: BBa_K1179002
start: 1
end: 1174
orientation: inline

BBa_J85600
type: SequenceAnnotation
persistent identity: annotation2353737
display id: annotation2353737
version: 1
top level: BBa_K1179002
location: https://synbiohub.org/public/igem/BBa_K1179002/annotation2353737/range2353737/1
component: BBa_J85600

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type: Range
persistent identity: range2353737
display id: range2353737
version: 1
direction: direction/reverse
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start: 1175
end: 1196
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BBa_K1179009
type: SequenceAnnotation
persistent identity: annotation2353738
display id: annotation2353738
version: 1
top level: BBa_K1179002
location: https://synbiohub.org/public/igem/BBa_K1179002/annotation2353738/range2353738/1
component: BBa_K1179009

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type: Range
persistent identity: range2353738
display id: range2353738
version: 1
direction: direction/reverse
top level: BBa_K1179002
start: 1197
end: 5141
orientation: inline

BBa_K1982006
type: ComponentDefinition
persistent identity: BBa K1982006
display id: BBa_K1982006
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: tCas9-Vp64(Prokaryotic)
creator: Zexu Li
created: 2016-09-11T11:00:00Z
modified: 2016-10-18T10:24:08Z
discontinued: false
dominant: false
group u list: _2449_
m user id: 30176
owner id: 30176
owning group id: 9
status: status/Planning
bookmark: false
mutable description: The CRY2/CIBN interaction is entirely genetically encoded. The binding reverses within minutes in the dark, allowing rapid shutoff of transcription by placing samples in the dark. This fusion protein is for use in LACE(light-activated CRISPR/Cas9 effector) system, and a tCas9 fused to its N terminus. To regulate DNA transcription by blue light, the system is based on CRY2/CIBN interaction in which a light-mediated protein interaction brings together two protein (tCas9 and an activation domain VP64) . If we remove the stimulation of blue light, dark reversion of CRY2 will dissociate the interaction with CIBN and shut off transcription. tCas9 can be tagged with transcriptional activators, and targeting these dCas9 fusion proteins to the promoter region results in robust transcription activation of downstream target genes. This tCas9-based activators is the case that tCas9 fused directly to a single transcriptional activator( VP64).
mutable notes: mutant EcoR 1 : TGTTGGGAATTc--->TGTTGGGAATTg
mutable provenance: tCas9 is from GE Share company. VP64 is synthetic construct.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Device
role: SO:0000804
sequence annotation: TAATAA
sequence annotation: Prokaryotic rbs
sequence annotation: SV40 NLS
sequence annotation: tCas9
sequence annotation: FLAG
sequence annotation: Linker
sequence annotation: ATG
sequence annotation: VP16
sequence annotation: HA
sequence: https://synbiohub.org/public/igem/BBa_K1982006_sequence/1

TAATAA
type: SequenceAnnotation
persistent identity: annotation2517252
display id: annotation2517252
version: 1
top level: BBa_K1982006
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517252/range2517252/1

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type: Range
persistent identity: range2517252
display id: range2517252
version: 1
direction: direction/forward
top level: BBa_K1982006
start: 4363
end: 4368
orientation: inline

Prokaryotic rbs
type: SequenceAnnotation
persistent identity: annotation2517241
display id: annotation2517241
version: 1
top level: BBa_K1982006
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517241/range2517241/1

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type: Range
persistent identity: range2517241
display id: range2517241
version: 1
direction: direction/forward
top level: BBa_K1982006
start: 1
end: 18
orientation: inline

SV40 NLS
type: SequenceAnnotation
persistent identity: annotation2517244
display id: annotation2517244
version: 1
top level: BBa_K1982006
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517244/range2517244/1

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type: Range
persistent identity: range2517244
display id: range2517244
version: 1
direction: direction/forward
top level: BBa_K1982006
start: 4123
end: 4143
orientation: inline

tCas9
type: SequenceAnnotation
persistent identity: annotation2517243
display id: annotation2517243
version: 1
top level: BBa_K1982006
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517243/range2517243/1

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type: Range
persistent identity: range2517243
display id: range2517243
version: 1
direction: direction/reverse
top level: BBa_K1982006
start: 22
end: 4122
orientation: inline

FLAG
type: SequenceAnnotation
persistent identity: annotation2517251
display id: annotation2517251
version: 1
top level: BBa_K1982006
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517251/range2517251/1

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type: Range
persistent identity: range2517251
display id: range2517251
version: 1
direction: direction/forward
top level: BBa_K1982006
start: 4339
end: 4362
orientation: inline

Linker
type: SequenceAnnotation
persistent identity: annotation2517249
display id: annotation2517249
version: 1
top level: BBa_K1982006
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517249/range2517249/1

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type: Range
persistent identity: range2517249
display id: range2517249
version: 1
direction: direction/forward
top level: BBa_K1982006
start: 4300
end: 4305
orientation: inline

ATG
type: SequenceAnnotation
persistent identity: annotation2517242
display id: annotation2517242
version: 1
top level: BBa_K1982006
role: SO:0000318
role: feature/start
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517242/range2517242/1

https://synbiohub.org/public/igem/BBa_K1982006/annotation2517242/range2517242/1
type: Range
persistent identity: range2517242
display id: range2517242
version: 1
direction: direction/forward
top level: BBa_K1982006
start: 19
end: 21
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517247
display id: annotation2517247
version: 1
top level: BBa_K1982006
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517247/range2517247/1

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type: Range
persistent identity: range2517247
display id: range2517247
version: 1
direction: direction/forward
top level: BBa_K1982006
start: 4222
end: 4260
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517248
display id: annotation2517248
version: 1
top level: BBa_K1982006
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517248/range2517248/1

https://synbiohub.org/public/igem/BBa_K1982006/annotation2517248/range2517248/1
type: Range
persistent identity: range2517248
display id: range2517248
version: 1
direction: direction/forward
top level: BBa_K1982006
start: 4261
end: 4299
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517245
display id: annotation2517245
version: 1
top level: BBa_K1982006
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517245/range2517245/1

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type: Range
persistent identity: range2517245
display id: range2517245
version: 1
direction: direction/forward
top level: BBa_K1982006
start: 4144
end: 4182
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517246
display id: annotation2517246
version: 1
top level: BBa_K1982006
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517246/range2517246/1

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type: Range
persistent identity: range2517246
display id: range2517246
version: 1
direction: direction/forward
top level: BBa_K1982006
start: 4183
end: 4221
orientation: inline

HA
type: SequenceAnnotation
persistent identity: annotation2517250
display id: annotation2517250
version: 1
top level: BBa_K1982006
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1982006/annotation2517250/range2517250/1

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type: Range
persistent identity: range2517250
display id: range2517250
version: 1
direction: direction/forward
top level: BBa_K1982006
start: 4306
end: 4332
orientation: inline

BBa_K1160000
type: ComponentDefinition
persistent identity: BBa K1160000
display id: BBa_K1160000
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: coding sequence of Cas9 from CRISPR system type II
creator: Huang Xingxu
created: 2013-09-12T11:00:00Z
modified: 2015-05-08T01:09:32Z
discontinued: false
dominant: false
experience: experience/None
group u list: _1472_
m user id: 0
owner id: 16689
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable description: CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system provide protection against mobile genetic element by guided specific DNA cleavage. Cas9 (CRISPR-associated protein 9) belongs to type II CRISPR system and contains HNH motif and three RuvC-like motifs, homologous to HNH and RuvC endonuclease, respectively. Cas9 has strong DNA cleavage activity, thus it showed potential use in gene editing. This part contains the coding sequence of cas9 from Streptococcus pyogenes (UniProtKB AC number:Q99ZW2). Some "redundant" sequence was added to ensure the Cas9's normal function or just for the convenience to detect it in cell. The cas9 protein must be guided by two RNAs, crRNA(CRISPR RNA) and tracrRNA(trans-activating crRNA), to complete its specific DNA cleavage function. We build another part-a chimeic RNA-that combines the feature two RNAs. For more details, refer to part BBa_K1160001. With this part, you can 1)directly use it to express cas9 protein in cell or 2)use T7 RNA polymerase to get cas9's mRNA, which can be further modified to prepare authentic mRNA that can be used for translation in vivo.
mutable notes: to be added
mutable provenance: Streptococcus pyogenes
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000155
role: partType/Plasmid
sequence annotation: Ca9 coding sequence
sequence annotation: CMV enhancer
sequence annotation: AmpR
sequence annotation: BSD
sequence annotation: bGH poly(A) signal
sequence annotation: f1 ori
sequence annotation: CMV promoter
sequence annotation: ori
sequence: https://synbiohub.org/public/igem/BBa_K1160000_sequence/1

Ca9 coding sequence
type: SequenceAnnotation
persistent identity: annotation2363681
display id: annotation2363681
version: 1
top level: BBa_K1160000
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K1160000/annotation2363681/range2363681/1

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type: Range
persistent identity: range2363681
display id: range2363681
version: 1
direction: direction/forward
top level: BBa_K1160000
start: 904
end: 5010
orientation: inline

CMV enhancer
type: SequenceAnnotation
persistent identity: annotation2363683
display id: annotation2363683
version: 1
top level: BBa_K1160000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1160000/annotation2363683/range2363683/1

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type: Range
persistent identity: range2363683
display id: range2363683
version: 1
direction: direction/forward
top level: BBa_K1160000
start: 235
end: 614
orientation: inline

AmpR
type: SequenceAnnotation
persistent identity: annotation2363688
display id: annotation2363688
version: 1
top level: BBa_K1160000
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K1160000/annotation2363688/range2363688/1

https://synbiohub.org/public/igem/BBa_K1160000/annotation2363688/range2363688/1
type: Range
persistent identity: range2363688
display id: range2363688
version: 1
direction: direction/forward
top level: BBa_K1160000
start: 8163
end: 9023
orientation: inline

BSD
type: SequenceAnnotation
persistent identity: annotation2363686
display id: annotation2363686
version: 1
top level: BBa_K1160000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1160000/annotation2363686/range2363686/1

https://synbiohub.org/public/igem/BBa_K1160000/annotation2363686/range2363686/1
type: Range
persistent identity: range2363686
display id: range2363686
version: 1
direction: direction/forward
top level: BBa_K1160000
start: 6276
end: 6644
orientation: inline

bGH poly(A) signal
type: SequenceAnnotation
persistent identity: annotation2363684
display id: annotation2363684
version: 1
top level: BBa_K1160000
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1160000/annotation2363684/range2363684/1

https://synbiohub.org/public/igem/BBa_K1160000/annotation2363684/range2363684/1
type: Range
persistent identity: range2363684
display id: range2363684
version: 1
direction: direction/forward
top level: BBa_K1160000
start: 5120
end: 5344
orientation: inline

f1 ori
type: SequenceAnnotation
persistent identity: annotation2363685
display id: annotation2363685
version: 1
top level: BBa_K1160000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1160000/annotation2363685/range2363685/1

https://synbiohub.org/public/igem/BBa_K1160000/annotation2363685/range2363685/1
type: Range
persistent identity: range2363685
display id: range2363685
version: 1
direction: direction/forward
top level: BBa_K1160000
start: 5390
end: 5818
orientation: inline

CMV promoter
type: SequenceAnnotation
persistent identity: annotation2363682
display id: annotation2363682
version: 1
top level: BBa_K1160000
role: SO:0000167
role: feature/promoter
location: https://synbiohub.org/public/igem/BBa_K1160000/annotation2363682/range2363682/1

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type: Range
persistent identity: range2363682
display id: range2363682
version: 1
direction: direction/forward
top level: BBa_K1160000
start: 615
end: 818
orientation: inline

ori
type: SequenceAnnotation
persistent identity: annotation2363687
display id: annotation2363687
version: 1
top level: BBa_K1160000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1160000/annotation2363687/range2363687/1

https://synbiohub.org/public/igem/BBa_K1160000/annotation2363687/range2363687/1
type: Range
persistent identity: range2363687
display id: range2363687
version: 1
direction: direction/forward
top level: BBa_K1160000
start: 7404
end: 7992
orientation: inline

BBa_K1559003
type: ComponentDefinition
persistent identity: BBa K1559003
display id: BBa_K1559003
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: CRISPR/Cas9 system with Anderson high-expression constitutive promoter
creator: Xiuqi (Rex) Xia
created: 2014-10-12T11:00:00Z
modified: 2015-05-08T01:10:53Z
discontinued: false
dominant: false
group u list: _1944_
m user id: 0
owner id: 23047
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: TODO
mutable notes: TODO
mutable provenance: TODO
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Generator
component: BBa_J23119
component: BBa_K1559002
component: BBa_B0034
sequence annotation: BBa_B0034
sequence annotation: BBa_J23119
sequence annotation: BBa_K1559002
sequence: https://synbiohub.org/public/igem/BBa_K1559003_sequence/1

BBa_J23119
type: Component
persistent identity: component2418638
display id: component2418638
version: 1
top level: BBa_K1559003
access: public
definition: https://synbiohub.org/public/igem/BBa_J23119/1

BBa_K1559002
type: Component
persistent identity: component2418641
display id: component2418641
version: 1
top level: BBa_K1559003
access: public
definition: BBa_K1559002

BBa_B0034
type: Component
persistent identity: component2418640
display id: component2418640
version: 1
top level: BBa_K1559003
access: public
definition: https://synbiohub.org/public/igem/BBa_B0034/1

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2418640
display id: annotation2418640
version: 1
top level: BBa_K1559003
location: https://synbiohub.org/public/igem/BBa_K1559003/annotation2418640/range2418640/1
component: BBa_B0034

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type: Range
persistent identity: range2418640
display id: range2418640
version: 1
direction: direction/reverse
top level: BBa_K1559003
start: 36
end: 47
orientation: inline

BBa_J23119
type: SequenceAnnotation
persistent identity: annotation2418638
display id: annotation2418638
version: 1
top level: BBa_K1559003
location: https://synbiohub.org/public/igem/BBa_K1559003/annotation2418638/range2418638/1
component: BBa_J23119

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type: Range
persistent identity: range2418638
display id: range2418638
version: 1
direction: direction/reverse
top level: BBa_K1559003
start: 1
end: 35
orientation: inline

BBa_K1559002
type: SequenceAnnotation
persistent identity: annotation2418641
display id: annotation2418641
version: 1
top level: BBa_K1559003
location: https://synbiohub.org/public/igem/BBa_K1559003/annotation2418641/range2418641/1
component: BBa_K1559002

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type: Range
persistent identity: range2418641
display id: range2418641
version: 1
direction: direction/reverse
top level: BBa_K1559003
start: 48
end: 5127
orientation: inline

BBa_K1559004
type: ComponentDefinition
persistent identity: BBa K1559004
display id: BBa_K1559004
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: CRISPR/Cas9 system with Anderson medium-expression constitutive promoter
creator: Xiuqi (Rex) Xia
created: 2014-10-12T11:00:00Z
modified: 2015-05-08T01:10:53Z
discontinued: false
dominant: false
group u list: _1944_
m user id: 0
owner id: 23047
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: TODO
mutable notes: TODO
mutable provenance: TODO
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Generator
component: BBa_K1559002
component: BBa_J23108
component: BBa_B0034
sequence annotation: BBa_J23108
sequence annotation: BBa_K1559002
sequence annotation: BBa_B0034
sequence: https://synbiohub.org/public/igem/BBa_K1559004_sequence/1

BBa_K1559002
type: Component
persistent identity: component2418645
display id: component2418645
version: 1
top level: BBa_K1559004
access: public
definition: BBa_K1559002

BBa_J23108
type: Component
persistent identity: component2418642
display id: component2418642
version: 1
top level: BBa_K1559004
access: public
definition: https://synbiohub.org/public/igem/BBa_J23108/1

BBa_B0034
type: Component
persistent identity: component2418644
display id: component2418644
version: 1
top level: BBa_K1559004
access: public
definition: https://synbiohub.org/public/igem/BBa_B0034/1

BBa_J23108
type: SequenceAnnotation
persistent identity: annotation2418642
display id: annotation2418642
version: 1
top level: BBa_K1559004
location: https://synbiohub.org/public/igem/BBa_K1559004/annotation2418642/range2418642/1
component: BBa_J23108

https://synbiohub.org/public/igem/BBa_K1559004/annotation2418642/range2418642/1
type: Range
persistent identity: range2418642
display id: range2418642
version: 1
direction: direction/reverse
top level: BBa_K1559004
start: 1
end: 35
orientation: inline

BBa_K1559002
type: SequenceAnnotation
persistent identity: annotation2418645
display id: annotation2418645
version: 1
top level: BBa_K1559004
location: https://synbiohub.org/public/igem/BBa_K1559004/annotation2418645/range2418645/1
component: BBa_K1559002

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type: Range
persistent identity: range2418645
display id: range2418645
version: 1
direction: direction/reverse
top level: BBa_K1559004
start: 48
end: 5127
orientation: inline

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2418644
display id: annotation2418644
version: 1
top level: BBa_K1559004
location: https://synbiohub.org/public/igem/BBa_K1559004/annotation2418644/range2418644/1
component: BBa_B0034

https://synbiohub.org/public/igem/BBa_K1559004/annotation2418644/range2418644/1
type: Range
persistent identity: range2418644
display id: range2418644
version: 1
direction: direction/reverse
top level: BBa_K1559004
start: 36
end: 47
orientation: inline

BBa_K1994044
type: ComponentDefinition
persistent identity: BBa K1994044
display id: BBa_K1994044
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: dCas9 Promoter
creator: Egheosa Ogbomo
created: 2016-10-13T11:00:00Z
modified: 2016-10-19T02:12:18Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2461_
m user id: 30870
owner id: 28134
owning group id: 9
status: status/Planning
bookmark: false
mutable description: sgRNA with PAM-Proximal complementary sequence 5
mutable notes: sgRNA with PAM-Proximal complementary sequence 5
mutable provenance: sgRNA with PAM-Proximal complementary sequence 5
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000167
role: partType/Regulatory
sequence annotation: dCas9 Promoter
sequence annotation: Spacer
sequence: https://synbiohub.org/public/igem/BBa_K1994044_sequence/1

dCas9 Promoter
type: SequenceAnnotation
persistent identity: annotation2522296
display id: annotation2522296
version: 1
top level: BBa_K1994044
role: SO:0000167
role: feature/promoter
location: https://synbiohub.org/public/igem/BBa_K1994044/annotation2522296/range2522296/1

https://synbiohub.org/public/igem/BBa_K1994044/annotation2522296/range2522296/1
type: Range
persistent identity: range2522296
display id: range2522296
version: 1
direction: direction/forward
top level: BBa_K1994044
start: 18
end: 48
orientation: inline

Spacer
type: SequenceAnnotation
persistent identity: annotation2522272
display id: annotation2522272
version: 1
top level: BBa_K1994044
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1994044/annotation2522272/range2522272/1

https://synbiohub.org/public/igem/BBa_K1994044/annotation2522272/range2522272/1
type: Range
persistent identity: range2522272
display id: range2522272
version: 1
direction: direction/forward
top level: BBa_K1994044
start: 1
end: 17
orientation: inline

BBa_K1982007
type: ComponentDefinition
persistent identity: BBa K1982007
display id: BBa_K1982007
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Eukaryotic tCAS9
creator: Zexu Li
created: 2016-10-07T11:00:00Z
modified: 2016-10-09T12:04:45Z
discontinued: false
dominant: false
group u list: _2449_
m user id: 30176
owner id: 30176
owning group id: 9
status: status/Planning
bookmark: false
mutable description: tCas9 is a standardized (RFC 10) protein.Interacting with a DNA-binding RNA and fused with different effector domains it can be used for specific gene regulation. Cas9 is the main protein of the CRISPR/Cas system II of Streptococcus pyogenes. CRISPR systems protect bacteria and archaea from phages by recognizing and cleaving of invading phage DNA.This recognition is based on Watson Crick base pairing between a short RNA, called crRNA, and the complementary DNA strand. A second RNA, called tracrRNA, connects crRNA and Cas9. These three parts together form a protein-RNA-DNA complex with the targeted DNA strand [1] Cas9 became of great interest for research concerning DNA targeting, because of its ability to recognize site specific DNA strands by a crRNA. At first the functionality of Cas9 was modified by exchanging aminoacids. As a result, Cas9 was able to introduce mutations within the genome of several organisms by causing double strand breaks [2][3]. Then, it was converted from a nuclease to a nickase introducing single strand breaks [4] and lately it was converted to an enzymatically inactive form, called dCas9 [5]. This tCas9 is standardized (RFC 10). It can be used as a DNA binding protein, that can be fused with different effectors in order to regulate gene expression.
mutable notes: mutant Pst 1 :gctacctgcaG to gctacctgcaC mutant Pst 1 :actatctgcaG to actatctgcaC
mutable provenance: plasmid from GE Share company. [1] Westra E.R., Swarts D.C., Staals R.H., Jore M.M., Brouns S.J., van der Oost J. (2012). The CRISPRs, they are a-changin': how prokaryotes generate adaptive immunity. Annu Rev Genet. 46, 311-39 [2] Mali P., Yang L., Esvelt K.M., Aach J., Guell M., DiCarlo J.E., Norville J.E., Church G.M. (2013). RNA-guided human genome engineering via Cas9. Science 339(6121), 823-6 [3] Jiang W., Bikard D., Cox D., Zhang F., Marraffini L.A. (2013). RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat Biotechnol. 31(3), 233-9 [4] Cong, L., Ran, F.A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P.D., Wu, X., Jiang, W., Marraffini, L.A., Zhang, F. (2013). Multiplex Genome Engineering Using CRISPR/Cas Systems. Science 339 (6121), 819-23 [5] Qi L.S., Larson M.H., Gilbert L.A., Doudna J.A., Weissman J.S., Arkin A.P., Lim W.A. (2013). Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. Cell 152(5), 1173-83
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: atg
sequence annotation: Eukaryotic rbs
sequence: https://synbiohub.org/public/igem/BBa_K1982007_sequence/1

atg
type: SequenceAnnotation
persistent identity: annotation2488564
display id: annotation2488564
version: 1
top level: BBa_K1982007
role: SO:0000318
role: feature/start
location: https://synbiohub.org/public/igem/BBa_K1982007/annotation2488564/range2488564/1

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type: Range
persistent identity: range2488564
display id: range2488564
version: 1
direction: direction/forward
top level: BBa_K1982007
start: 19
end: 21
orientation: inline

Eukaryotic rbs
type: SequenceAnnotation
persistent identity: annotation2488563
display id: annotation2488563
version: 1
top level: BBa_K1982007
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K1982007/annotation2488563/range2488563/1

https://synbiohub.org/public/igem/BBa_K1982007/annotation2488563/range2488563/1
type: Range
persistent identity: range2488563
display id: range2488563
version: 1
direction: direction/forward
top level: BBa_K1982007
start: 1
end: 18
orientation: inline

BBa_K1150050
type: ComponentDefinition
persistent identity: BBa K1150050
display id: BBa_K1150050
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Truncated CMV dCas9 Device #4
creator: Natalie Louis and Lisa Schmunk
created: 2013-09-16T11:00:00Z
modified: 2015-05-08T01:09:28Z
discontinued: false
dominant: false
experience: experience/None
group u list: _1462_
m user id: 0
owner id: 16627
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable description: .
mutable notes: .
mutable provenance: .
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Device
role: SO:0000804
component: BBa_K1150067
component: BBa_K1150010
component: BBa_K747096
component: BBa_K1150016
component: BBa_K1150012
sequence annotation: BBa_K1150067
sequence annotation: BBa_K747096
sequence annotation: BBa_K1150010
sequence annotation: BBa_K1150012
sequence annotation: BBa_K1150016
sequence: https://synbiohub.org/public/igem/BBa_K1150050_sequence/1

BBa_K1150067
type: Component
persistent identity: component2354585
display id: component2354585
version: 1
top level: BBa_K1150050
access: public
definition: https://synbiohub.org/public/igem/BBa_K1150067/1

BBa_K1150010
type: Component
persistent identity: component2354587
display id: component2354587
version: 1
top level: BBa_K1150050
access: public
definition: https://synbiohub.org/public/igem/BBa_K1150010/1

BBa_K1150010
type: Component
persistent identity: component2354584
display id: component2354584
version: 1
top level: BBa_K1150050
access: public
definition: https://synbiohub.org/public/igem/BBa_K1150010/1

BBa_K747096
type: Component
persistent identity: component2354580
display id: component2354580
version: 1
top level: BBa_K1150050
access: public
definition: https://synbiohub.org/public/igem/BBa_K747096/1

BBa_K1150016
type: Component
persistent identity: component2354582
display id: component2354582
version: 1
top level: BBa_K1150050
access: public
definition: https://synbiohub.org/public/igem/BBa_K1150016/1

BBa_K1150012
type: Component
persistent identity: component2354589
display id: component2354589
version: 1
top level: BBa_K1150050
access: public
definition: https://synbiohub.org/public/igem/BBa_K1150012/1

BBa_K1150067
type: SequenceAnnotation
persistent identity: annotation2354585
display id: annotation2354585
version: 1
top level: BBa_K1150050
location: https://synbiohub.org/public/igem/BBa_K1150050/annotation2354585/range2354585/1
component: BBa_K1150067

https://synbiohub.org/public/igem/BBa_K1150050/annotation2354585/range2354585/1
type: Range
persistent identity: range2354585
display id: range2354585
version: 1
direction: direction/reverse
top level: BBa_K1150050
start: 653
end: 3367
orientation: inline

BBa_K747096
type: SequenceAnnotation
persistent identity: annotation2354580
display id: annotation2354580
version: 1
top level: BBa_K1150050
location: https://synbiohub.org/public/igem/BBa_K1150050/annotation2354580/range2354580/1
component: BBa_K747096

https://synbiohub.org/public/igem/BBa_K1150050/annotation2354580/range2354580/1
type: Range
persistent identity: range2354580
display id: range2354580
version: 1
direction: direction/reverse
top level: BBa_K1150050
start: 1
end: 580
orientation: inline

BBa_K1150010
type: SequenceAnnotation
persistent identity: annotation2354584
display id: annotation2354584
version: 1
top level: BBa_K1150050
location: https://synbiohub.org/public/igem/BBa_K1150050/annotation2354584/range2354584/1
component: BBa_K1150010

https://synbiohub.org/public/igem/BBa_K1150050/annotation2354584/range2354584/1
type: Range
persistent identity: range2354584
display id: range2354584
version: 1
direction: direction/reverse
top level: BBa_K1150050
start: 624
end: 644
orientation: inline

BBa_K1150010
type: SequenceAnnotation
persistent identity: annotation2354587
display id: annotation2354587
version: 1
top level: BBa_K1150050
location: https://synbiohub.org/public/igem/BBa_K1150050/annotation2354587/range2354587/1
component: BBa_K1150010

https://synbiohub.org/public/igem/BBa_K1150050/annotation2354587/range2354587/1
type: Range
persistent identity: range2354587
display id: range2354587
version: 1
direction: direction/reverse
top level: BBa_K1150050
start: 3376
end: 3396
orientation: inline

BBa_K1150012
type: SequenceAnnotation
persistent identity: annotation2354589
display id: annotation2354589
version: 1
top level: BBa_K1150050
location: https://synbiohub.org/public/igem/BBa_K1150050/annotation2354589/range2354589/1
component: BBa_K1150012

https://synbiohub.org/public/igem/BBa_K1150050/annotation2354589/range2354589/1
type: Range
persistent identity: range2354589
display id: range2354589
version: 1
direction: direction/reverse
top level: BBa_K1150050
start: 3405
end: 3626
orientation: inline

BBa_K1150016
type: SequenceAnnotation
persistent identity: annotation2354582
display id: annotation2354582
version: 1
top level: BBa_K1150050
location: https://synbiohub.org/public/igem/BBa_K1150050/annotation2354582/range2354582/1
component: BBa_K1150016

https://synbiohub.org/public/igem/BBa_K1150050/annotation2354582/range2354582/1
type: Range
persistent identity: range2354582
display id: range2354582
version: 1
direction: direction/reverse
top level: BBa_K1150050
start: 589
end: 615
orientation: inline

TC
type: ComponentDefinition
persistent identity: BBa K1982000
display id: BBa_K1982000
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: tCas9-CIBN (Prokaryotic LACE system)
creator: Zexu Li
created: 2016-09-09T11:00:00Z
modified: 2016-10-18T10:12:41Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _2449_
m user id: 30176
owner id: 30176
owning group id: 9
status: status/Planning
bookmark: false
mutable description: tCas9 is a standardized (RFC 10) protein.Interacting with a DNA-binding RNA and fused with different effector domains it can be used for specific gene regulation. Cas9 is the main protein of the CRISPR/Cas system II of Streptococcus pyogenes. CRISPR systems protect bacteria and archaea from phages by recognizing and cleaving of invading phage DNA.This recognition is based on Watson Crick base pairing between a short RNA, called crRNA, and the complementary DNA strand. A second RNA, called tracrRNA, connects crRNA and Cas9. These three parts together form a protein-RNA-DNA complex with the targeted DNA strand [1] Cas9 became of great interest for research concerning DNA targeting, because of its ability to recognize site specific DNA strands by a crRNA. At first the functionality of Cas9 was modified by exchanging aminoacids. As a result, Cas9 was able to introduce mutations within the genome of several organisms by causing double strand breaks [2][3]. Then, it was converted from a nuclease to a nickase introducing single strand breaks [4] and lately it was converted to an enzymatically inactive form, called dCas9 [5]. This tCas9 is standardized (RFC 10). It can be used as a DNA binding protein, that can be fused with different effectors in order to regulate gene expression.
mutable notes: mutant Pst 1 :gctacctgcaG to gctacctgcaC mutant Pst 1 :actatctgcaG to actatctgcaC
mutable provenance: GE Share
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Device
role: SO:0000804
sequence annotation: Prokaryotic rbs
sequence annotation: CIBN
sequence annotation: ATG
sequence annotation: TAATAA
sequence annotation: tCas9
sequence annotation: HA
sequence: https://synbiohub.org/public/igem/BBa_K1982000_sequence/1

Prokaryotic rbs
type: SequenceAnnotation
persistent identity: annotation2517184
display id: annotation2517184
version: 1
top level: TC
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K1982000/annotation2517184/range2517184/1

https://synbiohub.org/public/igem/BBa_K1982000/annotation2517184/range2517184/1
type: Range
persistent identity: range2517184
display id: range2517184
version: 1
direction: direction/forward
top level: TC
start: 1
end: 18
orientation: inline

CIBN
type: SequenceAnnotation
persistent identity: annotation2517188
display id: annotation2517188
version: 1
top level: TC
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K1982000/annotation2517188/range2517188/1

https://synbiohub.org/public/igem/BBa_K1982000/annotation2517188/range2517188/1
type: Range
persistent identity: range2517188
display id: range2517188
version: 1
direction: direction/forward
top level: TC
start: 4123
end: 4668
orientation: inline

ATG
type: SequenceAnnotation
persistent identity: annotation2517185
display id: annotation2517185
version: 1
top level: TC
role: SO:0000318
role: feature/start
location: https://synbiohub.org/public/igem/BBa_K1982000/annotation2517185/range2517185/1

https://synbiohub.org/public/igem/BBa_K1982000/annotation2517185/range2517185/1
type: Range
persistent identity: range2517185
display id: range2517185
version: 1
direction: direction/forward
top level: TC
start: 19
end: 21
orientation: inline

TAATAA
type: SequenceAnnotation
persistent identity: annotation2517187
display id: annotation2517187
version: 1
top level: TC
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K1982000/annotation2517187/range2517187/1

https://synbiohub.org/public/igem/BBa_K1982000/annotation2517187/range2517187/1
type: Range
persistent identity: range2517187
display id: range2517187
version: 1
direction: direction/forward
top level: TC
start: 4726
end: 4731
orientation: inline

tCas9
type: SequenceAnnotation
persistent identity: annotation2517189
display id: annotation2517189
version: 1
top level: TC
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K1982000/annotation2517189/range2517189/1

https://synbiohub.org/public/igem/BBa_K1982000/annotation2517189/range2517189/1
type: Range
persistent identity: range2517189
display id: range2517189
version: 1
direction: direction/forward
top level: TC
start: 22
end: 4122
orientation: inline

HA
type: SequenceAnnotation
persistent identity: annotation2517186
display id: annotation2517186
version: 1
top level: TC
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1982000/annotation2517186/range2517186/1

https://synbiohub.org/public/igem/BBa_K1982000/annotation2517186/range2517186/1
type: Range
persistent identity: range2517186
display id: range2517186
version: 1
direction: direction/forward
top level: TC
start: 4669
end: 4725
orientation: inline

BBa_K1150017
type: ComponentDefinition
persistent identity: BBa K1150017
display id: BBa_K1150017
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: dCas9 with CMV promoter
creator: Freiburg 2013
created: 2013-09-15T11:00:00Z
modified: 2015-05-08T01:09:27Z
discontinued: false
dominant: false
experience: experience/None
group u list: _1462_
m user id: 0
owner id: 16627
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable description: ..
mutable notes: ..
mutable provenance: ..
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Device
role: SO:0000804
component: BBa_K1150010
component: BBa_K747096
component: BBa_K1150016
component: BBa_K1150012
component: BBa_K1150000
sequence annotation: BBa_K1150012
sequence annotation: BBa_K1150010
sequence annotation: BBa_K747096
sequence annotation: BBa_K1150016
sequence annotation: BBa_K1150000
sequence: https://synbiohub.org/public/igem/BBa_K1150017_sequence/1

BBa_K1150010
type: Component
persistent identity: component2354367
display id: component2354367
version: 1
top level: BBa_K1150017
access: public
definition: https://synbiohub.org/public/igem/BBa_K1150010/1

BBa_K747096
type: Component
persistent identity: component2354363
display id: component2354363
version: 1
top level: BBa_K1150017
access: public
definition: https://synbiohub.org/public/igem/BBa_K747096/1

BBa_K1150016
type: Component
persistent identity: component2354365
display id: component2354365
version: 1
top level: BBa_K1150017
access: public
definition: https://synbiohub.org/public/igem/BBa_K1150016/1

BBa_K1150012
type: Component
persistent identity: component2354373
display id: component2354373
version: 1
top level: BBa_K1150017
access: public
definition: https://synbiohub.org/public/igem/BBa_K1150012/1

BBa_K1150000
type: Component
persistent identity: component2354369
display id: component2354369
version: 1
top level: BBa_K1150017
access: public
definition: BBa_K1150000

BBa_K1150010
type: Component
persistent identity: component2354371
display id: component2354371
version: 1
top level: BBa_K1150017
access: public
definition: https://synbiohub.org/public/igem/BBa_K1150010/1

BBa_K1150012
type: SequenceAnnotation
persistent identity: annotation2354373
display id: annotation2354373
version: 1
top level: BBa_K1150017
location: https://synbiohub.org/public/igem/BBa_K1150017/annotation2354373/range2354373/1
component: BBa_K1150012

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type: Range
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BBa_K1150010
type: SequenceAnnotation
persistent identity: annotation2354367
display id: annotation2354367
version: 1
top level: BBa_K1150017
location: https://synbiohub.org/public/igem/BBa_K1150017/annotation2354367/range2354367/1
component: BBa_K1150010

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type: Range
persistent identity: range2354367
display id: range2354367
version: 1
direction: direction/reverse
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start: 624
end: 644
orientation: inline

BBa_K747096
type: SequenceAnnotation
persistent identity: annotation2354363
display id: annotation2354363
version: 1
top level: BBa_K1150017
location: https://synbiohub.org/public/igem/BBa_K1150017/annotation2354363/range2354363/1
component: BBa_K747096

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type: Range
persistent identity: range2354363
display id: range2354363
version: 1
direction: direction/reverse
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start: 1
end: 580
orientation: inline

BBa_K1150010
type: SequenceAnnotation
persistent identity: annotation2354371
display id: annotation2354371
version: 1
top level: BBa_K1150017
location: https://synbiohub.org/public/igem/BBa_K1150017/annotation2354371/range2354371/1
component: BBa_K1150010

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type: Range
persistent identity: range2354371
display id: range2354371
version: 1
direction: direction/reverse
top level: BBa_K1150017
start: 4762
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BBa_K1150016
type: SequenceAnnotation
persistent identity: annotation2354365
display id: annotation2354365
version: 1
top level: BBa_K1150017
location: https://synbiohub.org/public/igem/BBa_K1150017/annotation2354365/range2354365/1
component: BBa_K1150016

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type: Range
persistent identity: range2354365
display id: range2354365
version: 1
direction: direction/reverse
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start: 589
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BBa_K1150000
type: SequenceAnnotation
persistent identity: annotation2354369
display id: annotation2354369
version: 1
top level: BBa_K1150017
location: https://synbiohub.org/public/igem/BBa_K1150017/annotation2354369/range2354369/1
component: BBa_K1150000

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type: Range
persistent identity: range2354369
display id: range2354369
version: 1
direction: direction/reverse
top level: BBa_K1150017
start: 653
end: 4753
orientation: inline

BBa_K1559007
type: ComponentDefinition
persistent identity: BBa K1559007
display id: BBa_K1559007
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: CRISPR/Cas9 system with pLac inducible promoter
creator: Xiuqi (Rex) Xia
created: 2014-10-12T11:00:00Z
modified: 2015-05-08T01:10:53Z
discontinued: false
dominant: false
group u list: _1944_
m user id: 0
owner id: 23047
owning group id: 9
sample status: Not in stock
status: status/Unavailable
bookmark: false
mutable description: TODO
mutable notes: TODO
mutable provenance: TODO
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Generator
component: BBa_K1559002
component: BBa_B0034
component: BBa_R0011
sequence annotation: BBa_B0034
sequence annotation: BBa_R0011
sequence annotation: BBa_K1559002
sequence: https://synbiohub.org/public/igem/BBa_K1559007_sequence/1

BBa_K1559002
type: Component
persistent identity: component2418673
display id: component2418673
version: 1
top level: BBa_K1559007
access: public
definition: BBa_K1559002

BBa_B0034
type: Component
persistent identity: component2418672
display id: component2418672
version: 1
top level: BBa_K1559007
access: public
definition: https://synbiohub.org/public/igem/BBa_B0034/1

BBa_R0011
type: Component
persistent identity: component2418666
display id: component2418666
version: 1
top level: BBa_K1559007
access: public
definition: https://synbiohub.org/public/igem/BBa_R0011/1

BBa_B0034
type: SequenceAnnotation
persistent identity: annotation2418672
display id: annotation2418672
version: 1
top level: BBa_K1559007
location: https://synbiohub.org/public/igem/BBa_K1559007/annotation2418672/range2418672/1
component: BBa_B0034

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type: Range
persistent identity: range2418672
display id: range2418672
version: 1
direction: direction/reverse
top level: BBa_K1559007
start: 56
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orientation: inline

BBa_R0011
type: SequenceAnnotation
persistent identity: annotation2418666
display id: annotation2418666
version: 1
top level: BBa_K1559007
location: https://synbiohub.org/public/igem/BBa_K1559007/annotation2418666/range2418666/1
component: BBa_R0011

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type: Range
persistent identity: range2418666
display id: range2418666
version: 1
direction: direction/forward
top level: BBa_K1559007
start: 1
end: 54
orientation: inline

BBa_K1559002
type: SequenceAnnotation
persistent identity: annotation2418673
display id: annotation2418673
version: 1
top level: BBa_K1559007
location: https://synbiohub.org/public/igem/BBa_K1559007/annotation2418673/range2418673/1
component: BBa_K1559002

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type: Range
persistent identity: range2418673
display id: range2418673
version: 1
direction: direction/reverse
top level: BBa_K1559007
start: 68
end: 5147
orientation: inline

BBa_K1218011
type: ComponentDefinition
persistent identity: BBa K1218011
display id: BBa_K1218011
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Cas9
creator: Sophia Liang
created: 2013-08-28T11:00:00Z
modified: 2015-05-08T01:09:42Z
discontinued: false
dominant: false
experience: experience/None
group u list: _1532_
m user id: 0
owner id: 17300
owning group id: 9
sample status: In stock
status: status/Available
bookmark: false
mutable description: CRISPR-Cas is a bacterial immune system that remembers and targets foreign viral DNA by storing DNA sequences, or spacers, between clustered regularly interspaced short palindromic repeats (CRISPRs). RNA transcripts of the spacers are then used to sense homologous DNA, which is cleaved by CRISPR-associated (Cas) proteins. This part codes for the tracrRNA, Cas9 protein, and minimal CRISPR array of a type II CRISPR-Cas system. The CRISPR array includes two CRISPR repeats separated by a spacer with two BsaI sites, which allow for the easy cloning of new spacers.
mutable notes: Mutagenesis to remove internal EcoRI site
mutable provenance: Streptococcus pyogenes
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence: https://synbiohub.org/public/igem/BBa_K1218011_sequence/1

VP64
type: ComponentDefinition
persistent identity: BBa K1982012
display id: BBa_K1982012
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: VP64 transcription activitor
creator: Zexu Li
created: 2016-10-17T11:00:00Z
modified: 2016-10-18T09:58:31Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _2449_
m user id: 30176
owner id: 30176
owning group id: 9
status: status/Planning
bookmark: false
mutable description: {| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right" ! colspan="2" style="background:#FFBF00;"|VP64 |- |'''Function''' |transcription activator |- |'''RFC standard''' |[http://parts.igem.org/Help:Assembly_standard_10 RFC 10] |- |'''Backbone''' |pSB1C3 |- |'''Organism''' |''synthetic construct'' |- |'''Source''' |synthetic construct |- |'''Submitted by''' |[http://2016.igem.org/Team:NEU-China NEU-China 2016] |} Tetrameric VP16 transcription activator domain
mutable notes: synthetic construct
mutable provenance: synthetic construct
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000417
role: partType/Protein Domain
sequence annotation: VP16
sequence annotation: Linker
sequence annotation: TAATAA
sequence annotation: FLAG
sequence annotation: HA
sequence annotation: SV40 NLS
sequence: https://synbiohub.org/public/igem/BBa_K1982012_sequence/1

VP16
type: SequenceAnnotation
persistent identity: annotation2517152
display id: annotation2517152
version: 1
top level: VP64
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982012/annotation2517152/range2517152/1

https://synbiohub.org/public/igem/BBa_K1982012/annotation2517152/range2517152/1
type: Range
persistent identity: range2517152
display id: range2517152
version: 1
direction: direction/forward
top level: VP64
start: 103
end: 141
orientation: inline

Linker
type: SequenceAnnotation
persistent identity: annotation2517154
display id: annotation2517154
version: 1
top level: VP64
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982012/annotation2517154/range2517154/1

https://synbiohub.org/public/igem/BBa_K1982012/annotation2517154/range2517154/1
type: Range
persistent identity: range2517154
display id: range2517154
version: 1
direction: direction/forward
top level: VP64
start: 181
end: 186
orientation: inline

TAATAA
type: SequenceAnnotation
persistent identity: annotation2517157
display id: annotation2517157
version: 1
top level: VP64
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K1982012/annotation2517157/range2517157/1

https://synbiohub.org/public/igem/BBa_K1982012/annotation2517157/range2517157/1
type: Range
persistent identity: range2517157
display id: range2517157
version: 1
direction: direction/forward
top level: VP64
start: 241
end: 246
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517150
display id: annotation2517150
version: 1
top level: VP64
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982012/annotation2517150/range2517150/1

https://synbiohub.org/public/igem/BBa_K1982012/annotation2517150/range2517150/1
type: Range
persistent identity: range2517150
display id: range2517150
version: 1
direction: direction/forward
top level: VP64
start: 22
end: 63
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517151
display id: annotation2517151
version: 1
top level: VP64
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982012/annotation2517151/range2517151/1

https://synbiohub.org/public/igem/BBa_K1982012/annotation2517151/range2517151/1
type: Range
persistent identity: range2517151
display id: range2517151
version: 1
direction: direction/forward
top level: VP64
start: 64
end: 102
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517153
display id: annotation2517153
version: 1
top level: VP64
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982012/annotation2517153/range2517153/1

https://synbiohub.org/public/igem/BBa_K1982012/annotation2517153/range2517153/1
type: Range
persistent identity: range2517153
display id: range2517153
version: 1
direction: direction/forward
top level: VP64
start: 142
end: 180
orientation: inline

FLAG
type: SequenceAnnotation
persistent identity: annotation2517156
display id: annotation2517156
version: 1
top level: VP64
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1982012/annotation2517156/range2517156/1

https://synbiohub.org/public/igem/BBa_K1982012/annotation2517156/range2517156/1
type: Range
persistent identity: range2517156
display id: range2517156
version: 1
direction: direction/forward
top level: VP64
start: 217
end: 240
orientation: inline

HA
type: SequenceAnnotation
persistent identity: annotation2517155
display id: annotation2517155
version: 1
top level: VP64
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1982012/annotation2517155/range2517155/1

https://synbiohub.org/public/igem/BBa_K1982012/annotation2517155/range2517155/1
type: Range
persistent identity: range2517155
display id: range2517155
version: 1
direction: direction/forward
top level: VP64
start: 187
end: 213
orientation: inline

SV40 NLS
type: SequenceAnnotation
persistent identity: annotation2517149
display id: annotation2517149
version: 1
top level: VP64
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982012/annotation2517149/range2517149/1

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type: Range
persistent identity: range2517149
display id: range2517149
version: 1
direction: direction/forward
top level: VP64
start: 1
end: 21
orientation: inline

BBa_K1982004
type: ComponentDefinition
persistent identity: BBa K1982004
display id: BBa_K1982004
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: tCas9-CIBN (Prokaryotic LACE system)
creator: Zexu Li
created: 2016-09-11T11:00:00Z
modified: 2016-10-18T10:12:30Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _2449_
m user id: 30176
owner id: 30176
owning group id: 9
status: status/Planning
bookmark: false
mutable description: Optogenetic systems enable precise spatial and temporal control of cell behavior. A light-activated CRISPR/Cas9 effector (LACE) system that induces transcription of endogenous genes in the presence of blue light.This was accomplished by fusing the light-inducible heterodimerizing proteins CRY2 and CIB1 to a transactivation domain and the catalytically inactive tCas9, respectively. The versatile LACE system can be easily directed to new DNA sequences for the dynamic regulation of endogenous genes[1]. we fused the light-inducible heterodimerizering proteins CRY2 and CIB1 from Arabidopsis thaliana to the VP64 transactivation domain and C-terminus of tCas9.
mutable notes: The CRY2/CIBN interaction is entirely genetically encoded. The binding reverses within minutes in the dark, allowing rapid shutoff of transcription by placing samples in the dark. This fusion protein is for use in LACE(light-activated CRISPR/Cas9 effector) system, and a tCas9 fused to its N terminus. To regulate DNA transcription by blue light, the system is based on CRY2/CIBN interaction in which a light-mediated protein interaction brings together two protein (tCas9 and an activation domain VP64) . If we remove the stimulation of blue light, dark reversion of CRY2 will dissociate the interaction with CIBN and shut off transcription.
mutable provenance: CRY2 and CIB1 are extracted from Arabidopsis thaliana. tCas9 is from GE Share company. VP64 is synthetic construct.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Device
role: SO:0000804
sequence annotation: ATG
sequence annotation: tCas9
sequence annotation: Prokaryotic rbs
sequence annotation: TAATAA
sequence annotation: HA
sequence annotation: CIBN
sequence: https://synbiohub.org/public/igem/BBa_K1982004_sequence/1

ATG
type: SequenceAnnotation
persistent identity: annotation2483005
display id: annotation2483005
version: 1
top level: BBa_K1982004
role: SO:0000318
role: feature/start
location: https://synbiohub.org/public/igem/BBa_K1982004/annotation2483005/range2483005/1

https://synbiohub.org/public/igem/BBa_K1982004/annotation2483005/range2483005/1
type: Range
persistent identity: range2483005
display id: range2483005
version: 1
direction: direction/forward
top level: BBa_K1982004
start: 4105
end: 4107
orientation: inline

tCas9
type: SequenceAnnotation
persistent identity: annotation2483004
display id: annotation2483004
version: 1
top level: BBa_K1982004
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K1982004/annotation2483004/range2483004/1

https://synbiohub.org/public/igem/BBa_K1982004/annotation2483004/range2483004/1
type: Range
persistent identity: range2483004
display id: range2483004
version: 1
direction: direction/reverse
top level: BBa_K1982004
start: 22
end: 4122
orientation: inline

Prokaryotic rbs
type: SequenceAnnotation
persistent identity: annotation2517191
display id: annotation2517191
version: 1
top level: BBa_K1982004
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K1982004/annotation2517191/range2517191/1

https://synbiohub.org/public/igem/BBa_K1982004/annotation2517191/range2517191/1
type: Range
persistent identity: range2517191
display id: range2517191
version: 1
direction: direction/forward
top level: BBa_K1982004
start: 1
end: 18
orientation: inline

TAATAA
type: SequenceAnnotation
persistent identity: annotation2483006
display id: annotation2483006
version: 1
top level: BBa_K1982004
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K1982004/annotation2483006/range2483006/1

https://synbiohub.org/public/igem/BBa_K1982004/annotation2483006/range2483006/1
type: Range
persistent identity: range2483006
display id: range2483006
version: 1
direction: direction/forward
top level: BBa_K1982004
start: 4726
end: 4731
orientation: inline

HA
type: SequenceAnnotation
persistent identity: annotation2517190
display id: annotation2517190
version: 1
top level: BBa_K1982004
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1982004/annotation2517190/range2517190/1

https://synbiohub.org/public/igem/BBa_K1982004/annotation2517190/range2517190/1
type: Range
persistent identity: range2517190
display id: range2517190
version: 1
direction: direction/forward
top level: BBa_K1982004
start: 4669
end: 4725
orientation: inline

ATG
type: SequenceAnnotation
persistent identity: annotation2483003
display id: annotation2483003
version: 1
top level: BBa_K1982004
role: SO:0000318
role: feature/start
location: https://synbiohub.org/public/igem/BBa_K1982004/annotation2483003/range2483003/1

https://synbiohub.org/public/igem/BBa_K1982004/annotation2483003/range2483003/1
type: Range
persistent identity: range2483003
display id: range2483003
version: 1
direction: direction/forward
top level: BBa_K1982004
start: 19
end: 21
orientation: inline

CIBN
type: SequenceAnnotation
persistent identity: annotation2483007
display id: annotation2483007
version: 1
top level: BBa_K1982004
role: SO:0000316
role: feature/protein
location: https://synbiohub.org/public/igem/BBa_K1982004/annotation2483007/range2483007/1

https://synbiohub.org/public/igem/BBa_K1982004/annotation2483007/range2483007/1
type: Range
persistent identity: range2483007
display id: range2483007
version: 1
direction: direction/reverse
top level: BBa_K1982004
start: 4123
end: 4668
orientation: inline

BBa_K1982008
type: ComponentDefinition
persistent identity: BBa K1982008
display id: BBa_K1982008
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: tCas9-CIBN (Eukaryotic LACE system)
creator: Zexu Li
created: 2016-10-08T11:00:00Z
modified: 2016-10-18T10:49:42Z
discontinued: false
dominant: false
group u list: _2449_
m user id: 30176
owner id: 30176
owning group id: 9
status: status/Planning
bookmark: false
mutable description: The NEU-China iGEM team 2016 designed a fusion protein consisting of tCas9 and CIBN for sequence-specific transactivation of a desired target locus (more information). We used our double truncated tCas9 (BBa_K1982007) impaired in its cleavage activity and fused it to the CIBN (BBa_K1982003). An prokaryotic RBS sequence from the Community collection(BBa_B0034) fused to the beginning of tCas9-CIBN. For detection of expression the fusion protein was tagged with a HA-epitope coding sequence (BBa_K1150016) .Figure 1 illustrates the detailed design of the whole device. Figure 1: Construct design. tCas9 was fused to CIBN. The resulting fusion construct was flanked by RBS sequences and tagged by a HA epitope. The pBad/araC promoter and rrnB T1 terminator were chosen to control gene expression. Optogenetic systems enable precise spatial and temporal control of cell behavior. A light-activated CRISPR/Cas9 effector (LACE) system that induces transcription of endogenous genes in the presence of blue light.This was accomplished by fusing the light-inducible heterodimerizing proteins CRY2 and CIB1 to a transactivation domain and the catalytically inactive tCas9, respectively. The versatile LACE system can be easily directed to new DNA sequences for the dynamic regulation of endogenous genes[1]. we fused the light-inducible heterodimerizering proteins CRY2 and CIB1 from Arabidopsis thaliana to the VP64 transactivation domain and C-terminus of tCas9.
mutable notes: The CRY2/CIBN interaction is entirely genetically encoded. The binding reverses within minutes in the dark, allowing rapid shutoff of transcription by placing samples in the dark. This fusion protein is for use in LACE(light-activated CRISPR/Cas9 effector) system, and a tCas9 fused to its N terminus. To regulate DNA transcription by blue light, the system is based on CRY2/CIBN interaction in which a light-mediated protein interaction brings together two protein (tCas9 and an activation domain VP64) . If we remove the stimulation of blue light, dark reversion of CRY2 will dissociate the interaction with CIBN and shut off transcription.
mutable provenance: CRY2 and CIB1 are extracted from Arabidopsis thaliana. tCas9 is from GE Share company. VP64 is synthetic construct.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: ATG
sequence annotation: Eukaryotic rbs
sequence annotation: HA
sequence annotation: tCas9
sequence annotation: BBa_K1982003
sequence annotation: TAATAA
sequence: https://synbiohub.org/public/igem/BBa_K1982008_sequence/1

ATG
type: SequenceAnnotation
persistent identity: annotation2517276
display id: annotation2517276
version: 1
top level: BBa_K1982008
role: SO:0000318
role: feature/start
location: https://synbiohub.org/public/igem/BBa_K1982008/annotation2517276/range2517276/1

https://synbiohub.org/public/igem/BBa_K1982008/annotation2517276/range2517276/1
type: Range
persistent identity: range2517276
display id: range2517276
version: 1
direction: direction/forward
top level: BBa_K1982008
start: 19
end: 21
orientation: inline

Eukaryotic rbs
type: SequenceAnnotation
persistent identity: annotation2517274
display id: annotation2517274
version: 1
top level: BBa_K1982008
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K1982008/annotation2517274/range2517274/1

https://synbiohub.org/public/igem/BBa_K1982008/annotation2517274/range2517274/1
type: Range
persistent identity: range2517274
display id: range2517274
version: 1
direction: direction/forward
top level: BBa_K1982008
start: 1
end: 18
orientation: inline

HA
type: SequenceAnnotation
persistent identity: annotation2517279
display id: annotation2517279
version: 1
top level: BBa_K1982008
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1982008/annotation2517279/range2517279/1

https://synbiohub.org/public/igem/BBa_K1982008/annotation2517279/range2517279/1
type: Range
persistent identity: range2517279
display id: range2517279
version: 1
direction: direction/forward
top level: BBa_K1982008
start: 4669
end: 4725
orientation: inline

tCas9
type: SequenceAnnotation
persistent identity: annotation2517275
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BBa_K1982003
type: SequenceAnnotation
persistent identity: annotation2517277
display id: annotation2517277
version: 1
top level: BBa_K1982008
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K1982008/annotation2517277/range2517277/1

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persistent identity: range2517277
display id: range2517277
version: 1
direction: direction/forward
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start: 4123
end: 4668
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TAATAA
type: SequenceAnnotation
persistent identity: annotation2517278
display id: annotation2517278
version: 1
top level: BBa_K1982008
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K1982008/annotation2517278/range2517278/1

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persistent identity: range2517278
display id: range2517278
version: 1
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start: 4726
end: 4731
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BBa_K1982011
type: ComponentDefinition
persistent identity: BBa K1982011
display id: BBa_K1982011
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: tCas9-Vp64(Eukaryoticc)
creator: Zexu Li
created: 2016-10-08T11:00:00Z
modified: 2016-10-18T10:57:53Z
discontinued: false
dominant: false
group u list: _2449_
m user id: 30176
owner id: 30176
owning group id: 9
status: status/Planning
bookmark: false
mutable description: {| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right" ! colspan="2" style="background:#FFBF00;"|RBS-tCas9-VP64-HA-FLAG |- |'''Function''' |gene activation |- |'''Use in''' |Eukaryotic cells |- |'''RFC standard''' |[http://parts.igem.org/Help:Assembly_standard_10 RFC 10] |- |'''Backbone''' |pSB1C3 |- |'''Submitted by''' |[http://2016.igem.org/Team:NEU-China NEU-China 2016] |} The CRY2/CIBN interaction is entirely genetically encoded. The binding reverses within minutes in the dark, allowing rapid shutoff of transcription by placing samples in the dark. This fusion protein is for use in LACE(light-activated CRISPR/Cas9 effector) system, and a tCas9 fused to its N terminus. To regulate DNA transcription by blue light, the system is based on CRY2/CIBN interaction in which a light-mediated protein interaction brings together two protein (tCas9 and an activation domain VP64) . If we remove the stimulation of blue light, dark reversion of CRY2 will dissociate the interaction with CIBN and shut off transcription. tCas9 can be tagged with transcriptional activators, and targeting these dCas9 fusion proteins to the promoter region results in robust transcription activation of downstream target genes. This tCas9-based activators is the case that tCas9 fused directly to a single transcriptional activator( VP64). <!-- Add more about the biology of this part here ===Usage and Biology=== Protein data table for BioBrick <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_ ">BBa_ automatically created by the BioBrick-AutoAnnotator version 1.0 Nucleotide sequence in RFC 10: (underlined part encodes the protein) ATGGACAAG ... GACGACAAATAATAA ORF from nucleotide position 1 to 4344 (excluding stop-codon) Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below) 1 101 201 301 401 501 601 701 801 901 1001 1101 1201 1301 1401 MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYL…
mutable notes: mutant Pst 1 :gctacctgcaG to gctacctgcaC mutant Pst 1 :actatctgcaG to actatctgcaC
mutable provenance: tCas9 is from GE Share company. VP64 is synthetic construct.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: Eukaryotic rbs
sequence annotation: VP16
sequence annotation: SV40 NLS
sequence annotation: FLAG
sequence annotation: tCas9
sequence annotation: TAATAA
sequence annotation: ATG
sequence annotation: HA
sequence annotation: Linker
sequence: https://synbiohub.org/public/igem/BBa_K1982011_sequence/1

Eukaryotic rbs
type: SequenceAnnotation
persistent identity: annotation2517313
display id: annotation2517313
version: 1
top level: BBa_K1982011
role: feature/rbs
role: SO:0000139
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517313/range2517313/1

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type: Range
persistent identity: range2517313
display id: range2517313
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start: 1
end: 18
orientation: inline

VP16
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persistent identity: annotation2517318
display id: annotation2517318
version: 1
top level: BBa_K1982011
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517318/range2517318/1

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type: Range
persistent identity: range2517318
display id: range2517318
version: 1
direction: direction/forward
top level: BBa_K1982011
start: 4183
end: 4221
orientation: inline

SV40 NLS
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persistent identity: annotation2517316
display id: annotation2517316
version: 1
top level: BBa_K1982011
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517316/range2517316/1

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type: Range
persistent identity: range2517316
display id: range2517316
version: 1
direction: direction/forward
top level: BBa_K1982011
start: 4123
end: 4143
orientation: inline

FLAG
type: SequenceAnnotation
persistent identity: annotation2517323
display id: annotation2517323
version: 1
top level: BBa_K1982011
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517323/range2517323/1

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type: Range
persistent identity: range2517323
display id: range2517323
version: 1
direction: direction/forward
top level: BBa_K1982011
start: 4339
end: 4362
orientation: inline

VP16
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persistent identity: annotation2517317
display id: annotation2517317
version: 1
top level: BBa_K1982011
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517317/range2517317/1

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type: Range
persistent identity: range2517317
display id: range2517317
version: 1
direction: direction/forward
top level: BBa_K1982011
start: 4144
end: 4182
orientation: inline

tCas9
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persistent identity: annotation2517315
display id: annotation2517315
version: 1
top level: BBa_K1982011
role: SO:0000316
role: feature/protein
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type: Range
persistent identity: range2517315
display id: range2517315
version: 1
direction: direction/forward
top level: BBa_K1982011
start: 22
end: 4122
orientation: inline

TAATAA
type: SequenceAnnotation
persistent identity: annotation2517324
display id: annotation2517324
version: 1
top level: BBa_K1982011
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517324/range2517324/1

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type: Range
persistent identity: range2517324
display id: range2517324
version: 1
direction: direction/forward
top level: BBa_K1982011
start: 4363
end: 4368
orientation: inline

ATG
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persistent identity: annotation2517314
display id: annotation2517314
version: 1
top level: BBa_K1982011
role: SO:0000318
role: feature/start
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517314/range2517314/1

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type: Range
persistent identity: range2517314
display id: range2517314
version: 1
direction: direction/forward
top level: BBa_K1982011
start: 19
end: 21
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517320
display id: annotation2517320
version: 1
top level: BBa_K1982011
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517320/range2517320/1

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type: Range
persistent identity: range2517320
display id: range2517320
version: 1
direction: direction/forward
top level: BBa_K1982011
start: 4261
end: 4299
orientation: inline

HA
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persistent identity: annotation2517322
display id: annotation2517322
version: 1
top level: BBa_K1982011
role: SO:0000324
role: feature/tag
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517322/range2517322/1

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type: Range
persistent identity: range2517322
display id: range2517322
version: 1
direction: direction/forward
top level: BBa_K1982011
start: 4306
end: 4332
orientation: inline

Linker
type: SequenceAnnotation
persistent identity: annotation2517321
display id: annotation2517321
version: 1
top level: BBa_K1982011
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517321/range2517321/1

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type: Range
persistent identity: range2517321
display id: range2517321
version: 1
direction: direction/forward
top level: BBa_K1982011
start: 4300
end: 4305
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517319
display id: annotation2517319
version: 1
top level: BBa_K1982011
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517319/range2517319/1

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type: Range
persistent identity: range2517319
display id: range2517319
version: 1
direction: direction/forward
top level: BBa_K1982011
start: 4222
end: 4260
orientation: inline