BBa_K1486025
1
BBa_K1486025
ADH1 Terminator
2014-09-25T11:00:00Z
2015-05-08T01:10:42Z
To edit
To edit
false
false
_1866_
0
21042
9
Not in stock
false
To edit
false
EPFL iGem team 2014
annotation2418776
1
Adh1 terminator
range2418776
1
1
188
BBa_K1969004
1
BBa_K1969004
GAL1 promoter
2016-10-04T11:00:00Z
2016-10-08T08:45:50Z
Saccharomyces cerevisiae genomic sequence.
Transcription terminator for alcohol dehydrogenase 1.
false
false
_2436_
29963
29963
9
false
We chose to PCR amplify it out of Saccharomyces cerevisiae genomic DNA.
false
Yijie Lai
annotation2487974
1
GAL1 promoter
range2487974
1
1
442
BBa_K1969011
1
BBa_K1969011
Linker sequence between ADH1 terminator and two coding sequences downstream GAL1 promoter
2016-10-09T11:00:00Z
2016-10-12T06:33:57Z
It is designed artificially.
This is the linker sequence between ADH1 terminator and Nano-lantern(cAMP-1.6) (downstream GAL1 promoter).
false
false
_2436_
29963
29963
9
false
It is designed for homologous recombination with the backbone plasmid used to insert in the HIS3 site in yeast.
false
Yijie Lai
annotation2495427
1
Linker sequence between ADH1 terminator and Nano-lantern(cAMP-1.6) (downstream GAL1 promoter)
range2495427
1
1
20
BBa_K1969010
1
BBa_K1969010
Linker sequence between GAL1 promoter and Nano-lantern(cAMP-1.6)
2016-10-09T11:00:00Z
2016-10-12T06:38:15Z
It's designed artificially.
This is the linker sequence between GAL1 promoter and Nano-lantern(cAMP-1.6), it is left after the two functional part are ligated using recombinase.
false
false
_2436_
29963
29963
9
false
As we originally don't have the 4 standard restriction enzymes require for the biobrick assembly, we use homologous recombination to link two functional parts together.
false
Yijie Lai
annotation2489593
1
Linker sequence between GAL1 promoter and Nano-lantern(cAMP-1.6)
range2489593
1
1
10
BBa_K1969000
1
BBa_K1969000
Nano-lantern(cAMP-1.6)
2016-10-03T11:00:00Z
2016-10-05T10:23:22Z
It is a engineered sequence created by professor Nagai and we obtained it from addgene (Plasmid #61245).
Nano-lantern(cAMP-1.6) is a fusion protein designed by professor Nagai's group, It is a detector for changes in intracellular cAMP level.
The reporter mainly contains three parts: the enhanced yellow fluorescent protein, Venus△C10, the mutated Renilla luciferase, RLuc8△N3, and the cAMP-recognition motif of human EPAC1 with a Q270E mutation(Saito, Chang et al. 2012). The yellow flourescent part is used to increase the emitted photon number of RLuc8△N3 by a Forster resonance energy transfer (FRET) mechanism, in which the excited energy of the luminescent substrate, coelenterazine, bound to RLuc is efficiently transferred intermolecularly to the acceptor Venus△C10. The cAMP-recognition motif of human EPAC1 is flanked by two split parts of RLuc8△N3 based on the complementation of split luciferase (CSL) mechanism, thus the conformational change in cAMP-recognition motif reconstitutes the catalytic activity of the split-Luc in a target-molecule-dependent fashion. Upon binding with cAMP, the fusion protein, Nano-lantern(cAMP-1.6), showed the signal increase of about 130%.
We aimed to use the reporter to detect the cAMP surge upon stimulation by ligands of G-protein coupled receptors, so cloned Nano-lantern(cAMP-1.6) downstream three kinds of promoters and navigated its expression, localization and function in yeast Saccharomyces cerevisiae.
false
false
_2436_
29963
29963
9
false
As it contains a YFP, we don't need to add any tag to monitor its cellular localization.
false
Yijie Lai
annotation2487756
1
Nano-lantern(cAMP-1.6)
range2487756
1
1
2103
annotation2487757
1
ATG
range2487757
1
1
3
annotation2487758
1
TAA
range2487758
1
2101
2103
BBa_K1969013
1
BBa_K1969013
Nano-lantern(cAMP-1.6) regulated by GAL1 promoter
2016-10-09T11:00:00Z
2016-10-12T06:41:07Z
The expression of Nano-lantern(cAMP-1.6) may affect the sensing resolution of intracellular cAMP level, so creating expression cassettes with different expression level is critical for the project. This cassette is one of median expression level.
The sequence of Nano-lantern(cAMP-1.6) is cloned downstream the yeast GAL1 promoter and ended with an ADH1 terminator to generate a median expression cassette in yeast Saccharomyces cerevisiae.
false
false
_2436_
29963
29963
9
false
Use a inducible promoter GAL1 to mediate the expression of reporter protein Nano-lantern(cAMP-1.6).
false
Yijie Lai
component2495441
1
BBa_K1969004
component2495451
1
BBa_K1486025
component2495443
1
BBa_K1969010
component2495449
1
BBa_K1969011
component2495447
1
BBa_K1969000
annotation2495451
1
BBa_K1486025
range2495451
1
2576
2763
annotation2495449
1
BBa_K1969011
range2495449
1
2556
2575
annotation2495443
1
BBa_K1969010
range2495443
1
443
452
annotation2495441
1
BBa_K1969004
range2495441
1
1
442
annotation2495447
1
BBa_K1969000
range2495447
1
453
2555
BBa_K1969011_sequence
1
ggcgcgccacttctaaataa
BBa_K1486025_sequence
1
gcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagtatgaggtcgctcttattgaccacacctctaccgg
BBa_K1969004_sequence
1
cggattagaagccgccgagcgggcgacagccctccgacggaagactctcctccgtgcgtcctcgtcttcaccggtcgcgttcctgaaacgcagatgtgcctcgcgccgcactgctccgaacaataaagattctacaatactagcttttatggttatgaagaggaaaaattggcagtaacctggccccacaaaccttcaaattaacgaatcaaattaacaaccataggatgataatgcgattagttttttagccttatttctggggtaattaatcagcgaagcgatgatttttgatctattaacagatatataaatggaaaagctgcataaccactttaactaatactttcaacattttcagtttgtattacttcttattcaaatgtcataaaagtatcaacaaaaaattgttaatatacctctatactttaacgtcaaggag
BBa_K1969010_sequence
1
atgtcgtacg
BBa_K1969000_sequence
1
atgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaattaatttgcactactggaaaactacctgttccatggccaacacttgtcactactttaggttatggtcttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcacagcagacaaacaaaagaatggaatcaaagcaaacttcaaaattagacacaacattgaagatggaggtgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcctatcaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggggtaccaaggtgtacgaccccgagcagaggaagaggatgatcaccggcccccagtggtgggccaggtgcaagcagatgaacgtgctggacagcttcatcaactactacgacagcgagaagcacgccgagaacgccgtgatcttcctgcacggcaacgccactagcagctacctgtggaggcacgtggtgccccacatcgagcccgtggccaggtgcatcatccccgatctgatcggcatgggcaagagcggcaagagcggcaacggcagctacaggctgctggaccactacaagtacctgaccgcctggttcgagcttctgaacctgcccaagaagatcatcttcgtgggccacgactggggcgccgccctggccttccactacgcctacgagcaccaggacaggatcaaggccatcgtgcacatggagagcgtggtggacgtgatcgagagctgggacgagtggccagacatcgaggaggacatcgccctgatcaagagcgaggagggcgagaagatggtgctggagaacaacttcttcgtggagaccgtgctgcccagcaagatcatgagaaagctggagcccgaggagttcgccgcctacctggagcccttcaaggagaagggcgaggtgagaagacccaccctgagctggcccagagagatccccctggtgaagggcctcgaggggcctgacgccctgctcactgtggcacttcgaaagcccccaggtcagcgcacggatgaagagctggacctcatctttgaggagctgctgcacatcaaggctgtggcccacctctccaactcggtgaagcgagaattagcggctgttctgctctttgaaccacacagcaaggcagggaccgtgttgttcagccagggggacaagggcacttcgtggtacattatctggaagggatctgtcaacgtggtgacccatggcaaggggctggtgaccaccctgcatgagggagatgattttggagagctggctctggtgaatgatgcaccccgggcagccaccatcatcctgcgagaagacaactgtcatttcctgcgtgtggacaagcaggacttcaaccgtatcatcaaggatgtggaggcaaagaccatgcggctggaagaacatggcaaagtggtgctggtgctggagagagccgcatgcggcaagcccgacgtggtgcagatcgtgagaaactacaacgcctacctgagagccagcgacgacctgcccaagctgttcatcgagggtgaccccggcttcttcagcaacgccatcgtggagggcgccaagaagttccccaacaccgagttcgtgaaggtgaagggcctgcacttcctccaggaggacgcccccgacgagatgggcaagtacatcaagagcttcgtggagagagtgctgaagaacgagcagtaa
BBa_K1969013_sequence
1
cggattagaagccgccgagcgggcgacagccctccgacggaagactctcctccgtgcgtcctcgtcttcaccggtcgcgttcctgaaacgcagatgtgcctcgcgccgcactgctccgaacaataaagattctacaatactagcttttatggttatgaagaggaaaaattggcagtaacctggccccacaaaccttcaaattaacgaatcaaattaacaaccataggatgataatgcgattagttttttagccttatttctggggtaattaatcagcgaagcgatgatttttgatctattaacagatatataaatggaaaagctgcataaccactttaactaatactttcaacattttcagtttgtattacttcttattcaaatgtcataaaagtatcaacaaaaaattgttaatatacctctatactttaacgtcaaggagatgtcgtacgatgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaattaatttgcactactggaaaactacctgttccatggccaacacttgtcactactttaggttatggtcttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcacagcagacaaacaaaagaatggaatcaaagcaaacttcaaaattagacacaacattgaagatggaggtgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcctatcaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggggtaccaaggtgtacgaccccgagcagaggaagaggatgatcaccggcccccagtggtgggccaggtgcaagcagatgaacgtgctggacagcttcatcaactactacgacagcgagaagcacgccgagaacgccgtgatcttcctgcacggcaacgccactagcagctacctgtggaggcacgtggtgccccacatcgagcccgtggccaggtgcatcatccccgatctgatcggcatgggcaagagcggcaagagcggcaacggcagctacaggctgctggaccactacaagtacctgaccgcctggttcgagcttctgaacctgcccaagaagatcatcttcgtgggccacgactggggcgccgccctggccttccactacgcctacgagcaccaggacaggatcaaggccatcgtgcacatggagagcgtggtggacgtgatcgagagctgggacgagtggccagacatcgaggaggacatcgccctgatcaagagcgaggagggcgagaagatggtgctggagaacaacttcttcgtggagaccgtgctgcccagcaagatcatgagaaagctggagcccgaggagttcgccgcctacctggagcccttcaaggagaagggcgaggtgagaagacccaccctgagctggcccagagagatccccctggtgaagggcctcgaggggcctgacgccctgctcactgtggcacttcgaaagcccccaggtcagcgcacggatgaagagctggacctcatctttgaggagctgctgcacatcaaggctgtggcccacctctccaactcggtgaagcgagaattagcggctgttctgctctttgaaccacacagcaaggcagggaccgtgttgttcagccagggggacaagggcacttcgtggtacattatctggaagggatctgtcaacgtggtgacccatggcaaggggctggtgaccaccctgcatgagggagatgattttggagagctggctctggtgaatgatgcaccccgggcagccaccatcatcctgcgagaagacaactgtcatttcctgcgtgtggacaagcaggacttcaaccgtatcatcaaggatgtggaggcaaagaccatgcggctggaagaacatggcaaagtggtgctggtgctggagagagccgcatgcggcaagcccgacgtggtgcagatcgtgagaaactacaacgcctacctgagagccagcgacgacctgcccaagctgttcatcgagggtgaccccggcttcttcagcaacgccatcgtggagggcgccaagaagttccccaacaccgagttcgtgaaggtgaagggcctgcacttcctccaggaggacgcccccgacgagatgggcaagtacatcaagagcttcgtggagagagtgctgaagaacgagcagtaaggcgcgccacttctaaataagcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagtatgaggtcgctcttattgaccacacctctaccgg
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