BBa_K1982006
1
BBa_K1982006
tCas9-Vp64(Prokaryotic)
2016-09-11T11:00:00Z
2016-10-18T10:24:08Z
tCas9 is from GE Share company. VP64 is synthetic construct.
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).
false
false
_2449_
30176
30176
9
false
mutant EcoR 1 : TGTTGGGAATTc--->TGTTGGGAATTg
false
Zexu Li
annotation2517249
1
Linker
range2517249
1
4300
4305
annotation2517252
1
TAATAA
range2517252
1
4363
4368
annotation2517241
1
Prokaryotic rbs
range2517241
1
1
18
annotation2517244
1
SV40 NLS
range2517244
1
4123
4143
annotation2517242
1
ATG
range2517242
1
19
21
annotation2517243
1
tCas9
range2517243
1
22
4122
annotation2517251
1
FLAG
range2517251
1
4339
4362
annotation2517246
1
VP16
range2517246
1
4183
4221
annotation2517250
1
HA
range2517250
1
4306
4332
annotation2517247
1
VP16
range2517247
1
4222
4260
annotation2517248
1
VP16
range2517248
1
4261
4299
annotation2517245
1
VP16
range2517245
1
4144
4182
BBa_K1982006_sequence
1
aaagaggagaaatactagatggacaagaagtactccattggcctcgccatcggaacaaatagcgtgggctgggctgtcatcacagatgagtacaaggtgcctagcaagaaatttaaggtgctgggaaatacagacagacatagcatcaagaagaacctcattggcgctctcctgtttgactccggcgaaacagccgaagctaccagactcaagagaaccgctaggagaaggtacaccagaaggaaaaacaggatttgctacctgcacgaaattttttccaacgagatggccaaggtggacgattccttcttccataggctggaagagagcttcctcgtggaggaagacaagaaacacgagaggcatcctatttttggcaatattgtggatgaggtcgcctaccatgagaagtatcccacaatctatcatctgagaaaaaaactggtggatagcaccgacaaggccgatctcaggctcatttatctcgctctggctcacatgatcaagtttaggggccacttcctgatcgaaggcgacctgaatcccgacaactccgacgtggacaaactgttcatccagctcgtccagacctacaatcaactcttcgaggagaaccccatcaatgcttccggcgtggatgccaaggccatcctgagcgctaggctctccaagtccaggaggctggaaaatctgatcgcccaactccctggagagaagaagaacggcctgtttggcaatctgattgccctgagcctcggactcacccccaacttcaagagcaacttcgatctcgccgaagacgccaaactccaactgagcaaggatacctacgacgacgatctcgataatctcctcgcccagatcggcgatcaatatgccgacctctttctggccgccaaaaacctgagcgacgctattctgctcagcgacattctcagggtgaatacagaaatcacaaaagcccccctgtccgccagcatgatcaaaaggtacgatgaacaccatcaggacctcaccctgctgaaggctctggtcaggcagcaactccccgaaaagtacaaggagattttctttgatcagtccaagaatggatatgctggctatattgatggaggcgcctcccaggaggaattttataaattcatcaagcccattctcgaaaagatggacggaaccgaagagctgctggtcaaactcaatagggaggatctgctgaggaagcaaaggaccttcgacaatggcagcatcccccaccagatccacctcggcgaactccacgctatcctcaggaggcaggaagacttctaccctttcctgaaggataacagggagaaaatcgagaaaatcctgaccttcagaatcccctactacgtcggacctctcgccaggggcaattccagattcgcctggatgacaaggaagagcgaggaaacaatcacaccatggaacttcgaagaagtggtcgataagggcgccagcgcccagagcttcattgaaaggatgaccaactttgataagaacctgcccaatgagaaggtgctgcctaagcactccctgctgtatgagtatttcaccgtgtataatgagctgaccaaggtcaagtacgtcaccgagggaatgagaaagcctgcttttctctccggcgagcagaaaaaagccatcgtggacctgctgttcaagaccaacaggaaggtgaccgtcaagcaactcaaggaggactactttaagaagattgagtgctttgatagcgtggaaattagcggagtcgaggacaggttcaatgcctccctcggaacatatcacgacctgctgaaaatcatcaaagacaaagattttctggataacgaggagaatgaagacattctggaggacattgtcctcaccctgaccctgtttgaggacagagagatgattgaagagaggctgaaaacctatgcccacctgttcgacgacaaggtgatgaagcagctcaaaagaaggagatataccggctggggcagactgtccaggaagctgatcaacggcattagggacaagcagagcggcaagaccattctcgactttctcaagtccgacggattcgccaacagaaactttatgcaactgatccacgatgacagcctcacctttaaggaagatattcagaaggctcaggtcagcggccaaggcgattccctccatgagcacatcgctaatctggctggctcccctgctatcaaaaagggcatcctccagacagtcaaagtcgtcgatgagctggtcaaggtgatgggcaggcataaacccgagaacattgtgattgagatggctagggagaaccagaccacccagaaaggccagaaaaacagcagagaaagaatgaagaggatcgaggagggcatcaaagaactgggcagccaaatcctcaaggagcaccccgtcgaaaatacacaactccagaacgaaaaactctacctctactatctgcacaacggcagagacatgtacgtggaccaggaactggacatcaacaggctctccgattacgatgtggacgccatcgtccctcagtcctttctgaaagatgatagcatcgacaacaaggtgctgaccaggtccgacaagaataggggcaagagcgataatgtgccctccgaggaggtcgtcaaaaaaatgaaaaactactggagacaactcctcaacgctaagctcatcacccaaagaaagttcgacaatctgaccaaagccgagaggggcggcctctccgaactggacaaggccggcttcatcaaaaggcaattggtggaaaccaggcagattacaaagcatgtcgctcaaattctcgatagcaggatgaataccaaatatgacgagaacgacaagctgatcagagaggtcaaggtcatcacactcaagtccaagctcgtgagcgacttcagaaaagatttccaattttataaagtcagggagatcaacaattaccaccacgctcacgacgcttatctcaacgctgtcgtgggaaccgccctgatcaaaaaataccccaagctggaaagcgagttcgtgtatggcgattataaagtgtacgacgtgaggaagatgatcgctaaaagcgagcaggaaatcggcaaggctacagccaagtactttttctactccaacattatgaacttcttcaagaccgagattaccctcgccaacggcgaaattaggaagaggcccctgattgaaacaaatggagaaacaggcgaaatcgtctgggacaagggcagggacttcgccacagtcagaaaagtgctgtccatgcctcaagtcaacatcgtcaaaaagaccgaggtgcagaccggcggctttagcaaagaaagcatcctgcccaagagaaactccgacaagctcatcgctaggaagaaggactgggaccctaagaaatacggaggatttgactcccctaccgtcgcctattccgtcctcgtcgtcgctaaggtggagaagggcaagagcaagaagctcaagagcgtcaaggagctgctgggaatcaccatcatggagaggagctccttcgaaaaaaaccctattgatttcctggaggccaagggctacaaggaggtcaagaaggacctcatcatcaagctgcccaaatacagcctcttcgaactggaaaatggcaggaagagaatgctcgctagggccggcgagctccagaaaggaaatgagctggctctgcccagcaagtacgtcaacttcctctatctcgccagccactatgaaaagctcaagggcagccccgaagacaatgagcagaagcagctcttcgtcgagcagcacaagcactacctcgatgaaatcatcgagcaaatcagcgagttttccaaaagggtgattctcgccgacgctaacctcgataaggtcctctccgcttacaacaagcatagagacaagcccatcagagaacaggccgagaacatcatccacctgtttacactcacaaacctcggagcccctgccgcttttaaatacttcgatacaaccattgataggaagaggtacacctccaccaaggaggtgctggatgctaccctgattcatcaatccatcacaggactctacgaaacaaggattgacctgtcccaactgggaggcgacccgaaaaagaaacgcaaagttgggcgcgccgacgcgctggacgatttcgatctcgacatgctgggttctgatgccctcgatgactttgacctggatatgttgggaagcgacgcattggatgactttgatctggacatgctcggctccgatgctctggacgatttcgatctcgatatgttaattaactacccgtacgacgttccggactacgcttctgactacaaggacgacgacgacaaataataa
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
James Alastair McLaughlin
Chris J. Myers
2017-03-06T15:00:00.000Z