BBa_K1982011
type: ComponentDefinition
persistent identity: BBa K1982011
display id: BBa_K1982011
version: 1
was generated by: iGEM to SBOL conversion
description: tCas9-Vp64(Eukaryoticc)
created: 2016-10-08T11:00:00Z
modified: 2016-10-18T10:57:53Z
mutable provenance: 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-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…
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: mutant Pst 1 :gctacctgcaG to gctacctgcaC mutant Pst 1 :actatctgcaG to actatctgcaC
star: false
creator: Zexu Li
type: DnaRegion
role: partType/Coding
role: SO:0000316
sequence annotation: Eukaryotic rbs
sequence annotation: ATG
sequence annotation: tCas9
sequence annotation: SV40 NLS
sequence annotation: VP16
sequence annotation: Linker
sequence annotation: HA
sequence annotation: FLAG
sequence annotation: TAATAA
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
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517313/range2517313/1
role: SO:0000139
role: feature/rbs

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517313/range2517313/1
type: Range
persistent identity: range2517313
display id: range2517313
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 1
end: 18
orientation: inline

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

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517314/range2517314/1
type: Range
persistent identity: range2517314
display id: range2517314
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 19
end: 21
orientation: inline

tCas9
type: SequenceAnnotation
persistent identity: annotation2517315
display id: annotation2517315
version: 1
top level: BBa_K1982011
location: https://synbiohub.org/public/igem/BBa_K1982011/annotation2517315/range2517315/1
role: SO:0000316
role: feature/protein

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517315/range2517315/1
type: Range
persistent identity: range2517315
display id: range2517315
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 22
end: 4122
orientation: inline

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

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517316/range2517316/1
type: Range
persistent identity: range2517316
display id: range2517316
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 4123
end: 4143
orientation: inline

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

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517317/range2517317/1
type: Range
persistent identity: range2517317
display id: range2517317
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 4144
end: 4182
orientation: inline

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

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517318/range2517318/1
type: Range
persistent identity: range2517318
display id: range2517318
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 4183
end: 4221
orientation: inline

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

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517319/range2517319/1
type: Range
persistent identity: range2517319
display id: range2517319
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 4222
end: 4260
orientation: inline

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

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517320/range2517320/1
type: Range
persistent identity: range2517320
display id: range2517320
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 4261
end: 4299
orientation: inline

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

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517321/range2517321/1
type: Range
persistent identity: range2517321
display id: range2517321
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 4300
end: 4305
orientation: inline

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

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517322/range2517322/1
type: Range
persistent identity: range2517322
display id: range2517322
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 4306
end: 4332
orientation: inline

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

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517323/range2517323/1
type: Range
persistent identity: range2517323
display id: range2517323
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 4339
end: 4362
orientation: inline

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

https://synbiohub.org/public/igem/BBa_K1982011/annotation2517324/range2517324/1
type: Range
persistent identity: range2517324
display id: range2517324
version: 1
top level: BBa_K1982011
direction: direction/forward
start: 4363
end: 4368
orientation: inline

https://synbiohub.org/public/igem/BBa_K1982011_sequence/1
type: Sequence
persistent identity: BBa K1982011 sequence
display id: BBa_K1982011_sequence
version: 1
was generated by: iGEM to SBOL conversion
owned by: james
owned by: myers
elements: cccgccgccaccatggagatggacaagaagtactccattggcctcgccatcggaacaaatagcgtgggctgggctgtcatcacagatgagtacaaggtgcctagcaagaaatttaaggtgctgggaaatacagacagacatagcatcaagaagaacctcattggcgctctcctgtttgactccggcgaaacagccgaagctaccagactcaagagaaccgctaggagaaggtacaccagaaggaaaaacaggatttgctacctgcacgaaattttttccaacgagatggccaaggtggacgattccttcttccataggctggaagagagcttcctcgtggaggaagacaagaaacacgagaggcatcctatttttggcaatattgtggatgaggtcgcctaccatgagaagtatcccacaatctatcatctgagaaaaaaactggtggatagcaccgacaaggccgatctcaggctcatttatctcgctctggctcacatgatcaagtttaggggccacttcctgatcgaaggcgacctgaatcccgacaactccgacgtggacaaactgttcatccagctcgtccagacctacaatcaactcttcgaggagaaccccatcaatgcttccggcgtggatgccaaggccatcctgagcgctaggctctccaagtccaggaggctggaaaatctgatcgcccaactccctggagagaagaagaacggcctgtttggcaatctgattgccctgagcctcggactcacccccaacttcaagagcaacttcgatctcgccgaagacgccaaactccaactgagcaaggatacctacgacgacgatctcgataatctcctcgcccagatcggcgatcaatatgccgacctctttctggccgccaaaaacctgagcgacgctattctgctcagcgacattctcagggtgaatacagaaatcacaaaagcccccctgtccgccagcatgatcaaaaggtacgatgaacaccatcaggacctcaccctgctgaaggctctggtcaggcagcaactccccgaaaagtacaaggagattttctttgatcagtccaagaatggatatgctggctatattgatggaggcgcctcccaggaggaattttataaattcatcaagcccattctcgaaaagatggacggaaccgaagagctgctggtcaaactcaatagggaggatctgctgaggaagcaaaggaccttcgacaatggcagcatcccccaccagatccacctcggcgaactccacgctatcctcaggaggcaggaagacttctaccctttcctgaaggataacagggagaaaatcgagaaaatcctgaccttcagaatcccctactacgtcggacctctcgccaggggcaattccagattcgcctggatgacaaggaagagcgaggaaacaatcacaccatggaacttcgaagaagtggtcgataagggcgccagcgcccagagcttcattgaaaggatgaccaactttgataagaacctgcccaatgagaaggtgctgcctaagcactccctgctgtatgagtatttcaccgtgtataatgagctgaccaaggtcaagtacgtcaccgagggaatgagaaagcctgcttttctctccggcgagcagaaaaaagccatcgtggacctgctgttcaagaccaacaggaaggtgaccgtcaagcaactcaaggaggactactttaagaagattgagtgctttgatagcgtggaaattagcggagtcgaggacaggttcaatgcctccctcggaacatatcacgacctgctgaaaatcatcaaagacaaagattttctggataacgaggagaatgaagacattctggaggacattgtcctcaccctgaccctgtttgaggacagagagatgattgaagagaggctgaaaacctatgcccacctgttcgacgacaaggtgatgaagcagctcaaaagaaggagatataccggctggggcag…
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