BBa_K1982010
type: ComponentDefinition
persistent identity: BBa K1982010
display id: BBa_K1982010
version: 1
was generated by: iGEM to SBOL conversion
description: CRY2-VP64(Eukaryotic LACE system)
created: 2016-10-08T11:00:00Z
modified: 2016-10-18T10:53:31Z
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
mutable description: {| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right" ! colspan="2" style="background:#FFBF00;"|RBS-CRY2-VP64-HA-FLAG |- |'''Function''' |gene activation |- |'''Use in''' |Prokaryotic 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 VP64 fused to its C 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. We used the full-length CRY2 (BBa_K1982002) and fused it to the transcription activator domain VP64 (BBa_K1982000). An prokaryotic RBS sequence from the Community collection(BBa_B0034) fused to the beginning of CRY2-VP64. For detection of expression the fusion protein was tagged with a FLAG-epitope coding sequence (gactacaaggacgacgacgacaaa) .Figure 1 illustrates the detailed design of the whole device. <!-- 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) ATGAAGATG ... GACGACAAATAATAA ORF from nucleotide position 1 to 2076 (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 MKMDKKTIVWFRRDL…
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 notes: no
star: false
creator: Zexu Li
type: DnaRegion
role: SO:0000316
role: partType/Coding
sequence annotation: Eukaryotic rbs
sequence annotation: ATG
sequence annotation: CRY2
sequence annotation: SV40 NLS
sequence annotation: VP16
sequence annotation: Linker
sequence annotation: HA
sequence annotation: FLAG
sequence annotation: TAA
sequence: https://synbiohub.org/public/igem/BBa_K1982010_sequence/1

Eukaryotic rbs
type: SequenceAnnotation
persistent identity: annotation2517280
display id: annotation2517280
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517280/range2517280/1
role: SO:0000139
role: feature/rbs

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type: Range
persistent identity: range2517280
display id: range2517280
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 1
end: 18
orientation: inline

ATG
type: SequenceAnnotation
persistent identity: annotation2517281
display id: annotation2517281
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517281/range2517281/1
role: feature/start
role: SO:0000318

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type: Range
persistent identity: range2517281
display id: range2517281
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 19
end: 21
orientation: inline

CRY2
type: SequenceAnnotation
persistent identity: annotation2517282
display id: annotation2517282
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517282/range2517282/1
role: feature/protein
role: SO:0000316

https://synbiohub.org/public/igem/BBa_K1982010/annotation2517282/range2517282/1
type: Range
persistent identity: range2517282
display id: range2517282
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 22
end: 1854
orientation: inline

SV40 NLS
type: SequenceAnnotation
persistent identity: annotation2517283
display id: annotation2517283
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517283/range2517283/1
role: feature/misc
role: SO:0000110

https://synbiohub.org/public/igem/BBa_K1982010/annotation2517283/range2517283/1
type: Range
persistent identity: range2517283
display id: range2517283
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 1855
end: 1875
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517284
display id: annotation2517284
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517284/range2517284/1
role: SO:0000110
role: feature/misc

https://synbiohub.org/public/igem/BBa_K1982010/annotation2517284/range2517284/1
type: Range
persistent identity: range2517284
display id: range2517284
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 1876
end: 1914
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517285
display id: annotation2517285
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517285/range2517285/1
role: SO:0000110
role: feature/misc

https://synbiohub.org/public/igem/BBa_K1982010/annotation2517285/range2517285/1
type: Range
persistent identity: range2517285
display id: range2517285
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 1915
end: 1953
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517286
display id: annotation2517286
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517286/range2517286/1
role: SO:0000110
role: feature/misc

https://synbiohub.org/public/igem/BBa_K1982010/annotation2517286/range2517286/1
type: Range
persistent identity: range2517286
display id: range2517286
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 1954
end: 1992
orientation: inline

VP16
type: SequenceAnnotation
persistent identity: annotation2517287
display id: annotation2517287
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517287/range2517287/1
role: feature/misc
role: SO:0000110

https://synbiohub.org/public/igem/BBa_K1982010/annotation2517287/range2517287/1
type: Range
persistent identity: range2517287
display id: range2517287
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 1993
end: 2031
orientation: inline

Linker
type: SequenceAnnotation
persistent identity: annotation2517288
display id: annotation2517288
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517288/range2517288/1
role: SO:0000110
role: feature/misc

https://synbiohub.org/public/igem/BBa_K1982010/annotation2517288/range2517288/1
type: Range
persistent identity: range2517288
display id: range2517288
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 2032
end: 2037
orientation: inline

HA
type: SequenceAnnotation
persistent identity: annotation2517289
display id: annotation2517289
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517289/range2517289/1
role: feature/tag
role: SO:0000324

https://synbiohub.org/public/igem/BBa_K1982010/annotation2517289/range2517289/1
type: Range
persistent identity: range2517289
display id: range2517289
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 2037
end: 2063
orientation: inline

FLAG
type: SequenceAnnotation
persistent identity: annotation2517290
display id: annotation2517290
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517290/range2517290/1
role: feature/tag
role: SO:0000324

https://synbiohub.org/public/igem/BBa_K1982010/annotation2517290/range2517290/1
type: Range
persistent identity: range2517290
display id: range2517290
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 2071
end: 2094
orientation: inline

TAA
type: SequenceAnnotation
persistent identity: annotation2517291
display id: annotation2517291
version: 1
top level: BBa_K1982010
location: https://synbiohub.org/public/igem/BBa_K1982010/annotation2517291/range2517291/1
role: feature/stop
role: SO:0000319

https://synbiohub.org/public/igem/BBa_K1982010/annotation2517291/range2517291/1
type: Range
persistent identity: range2517291
display id: range2517291
version: 1
top level: BBa_K1982010
direction: direction/forward
start: 2095
end: 2100
orientation: inline

https://synbiohub.org/public/igem/BBa_K1982010_sequence/1
type: Sequence
persistent identity: BBa K1982010 sequence
display id: BBa_K1982010_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: cccgccgccaccatggagatgaagatggacaaaaagactatagtttggtttagaagagacctaaggattgaggataatcctgcattagcagcagctgctcacgaaggatctgtttttcctgtcttcatttggtgtcctgaagaagaaggacagttttatcctggaagagcttcaagatggtggatgaaacaatcacttgctcacttatctcaatccttgaaggctcttggatctgacctcactttaatcaaaacccacaacacgatttcagcgatcttggattgtatccgcgttaccggtgctacaaaagtcgtctttaaccacctctatgatcctgtttcgttagttcgggaccataccgtaaaggagaagctggtggaacgtgggatctctgtgcaaagctacaatggagatctattgtatgaaccgtgggagatatactgcgaaaagggcaaaccttttacgagtttcaattcttactggaagaaatgcttagatatgtcgattgaatccgttatgcttcctcctccttggcggttgatgccaataactgctgcggctgaagcgatttgggcgtgttcgattgaagaactagggctggagaatgaggccgagaaaccgagcaatgcgttgttaactagagcttggtctccaggatggagcaatgctgataagttactaaatgagttcatcgagaagcagttgatagattatgcaaagaacagcaagaaagttgttgggaattgtacttcactactttctccgtatctccatttcggggaaataagcgtcagacacgttttccagtgtgcccggatgaaacaaattatatgggcaagagataagaacagtgaaggagaagaaagtgcagatctttttcttaggggaatcggtttaagagagtattctcggtatatatgtttcaacttcccgtttactcacgagcaatcgttgttgagtcatcttcggtttttcccttgggatgctgatgttgataagttcaaggcctggagacaaggcaggaccggttatccgttggtggatgccggaatgagagagctttgggctaccggatggatgcataacagaataagagtgattgtttcaagctttgctgtgaagtttcttctccttccatggaaatggggaatgaagtatttctgggatacacttttggatgctgatttggaatgtgacatccttggctggcagtatatctctgggagtatccccgatggccacgagcttgatcgcttggacaatcccgcgttacaaggcgccaaatatgacccagaaggtgagtacataaggcaatggcttcccgagcttgcgagattgccaactgaatggatccatcatccatgggacgctcctttaaccgtactcaaagcttctggtgtggaactcggaacaaactatgcgaaacccattgtagacatcgacacagctcgtgagctactagctaaagctatttcaagaacccgtgaagcacagatcatgatcggagcagcacctgatgagattgtagcagatagcttcgaggccttaggggctaataccattaaagaacctggtctttgcccatctgtgtcttctaatgaccaacaagtaccttcggctgttcgttacaacgggtcaaagagagtgaaacctgaggaagaagaagagagagacatgaagaaatctaggggattcgatgaaagggagttgttttcgactgctgaatcttcttcttcttcgagtgtgtttttcgtttcgcagtcttgctcgttggcatcagaagggaagaatctggaaggtattcaagattcatctgatcagattactacaagtttgggaaaaaatggttgcaaaccgaaaaagaaacgcaaagttgggcgcgccgacgcgctggacgatttcgatctcgacatgctgggttctgatgccctcgatgactttgacctggatatgttgggaagcgacgcattggatgactttgatctggacatgctcggctc…
encoding: naseq.html

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