BBa_K801012
1
BBa_K801012
ADH1 yeast terminator
2012-09-20T11:00:00Z
2015-05-08T01:13:24Z
Extracted from genomic DNA of ''S. cerevisiae'' using PCR.
Released HQ 2013
ADH1 terminator of ''S. cerevisiae''
false
false
_1057_
0
13624
9
In stock
false
-
false
Georg Sch??tzinger
annotation2202145
1
ADH1-Terminator
range2202145
1
1
349
BBa_K530008
1
BBa_K530008
TDH3 Yeast Promoter
2011-08-14T11:00:00Z
2015-05-08T01:12:35Z
http://www.yeastgenome.org/cgi-bin/locus.fpl?locus=TDH3
Released HQ 2013
TDH3 Promoter is used to regulate the expression of genes within the genome of Saccharomyces Cerevisiae or baker's yeast.
false
false
_696_
0
8447
9
In stock
true
Promoter was extracted from purified genomic DNA from Saccharomyces Cerevisiae. The PCR reaction performed also served to add the biobrick prefix and suffix to the promoter.
false
Daniel Wolozny
annotation2125804
1
TDH3 Promoter
range2125804
1
1
500
BBa_K1462970
1
BBa_K1462970
TDH3+CTS1-2+GFP+ADH1
2014-10-14T11:00:00Z
2015-05-08T01:10:36Z
The targetting sequence derived from yeast can be located at cytoderm.
A COOH-terminal 12-amino acid serve as a typical cleavable signal sequence.
false
false
_1841_
0
21372
9
Not in stock
true
We use a constitutive promoter and easily express
false
LinZhou Li,YaRan Zhao
component2419981
1
BBa_K1462950
component2419973
1
BBa_K530008
component2419983
1
BBa_K801012
annotation2419973
1
BBa_K530008
range2419973
1
1
500
annotation2419981
1
BBa_K1462950
range2419981
1
507
1331
annotation2419983
1
BBa_K801012
range2419983
1
1340
1688
BBa_K1462930
1
BBa_K1462930
CTS1-1
2014-10-14T11:00:00Z
2015-05-08T01:10:36Z
The targetting sequence derived from yeast can be located at cytoderm.
CTS
false
false
_1841_
0
21372
9
Not in stock
false
cut short the length of the sequence.
false
LinZhou Li,YaRan Zhao
BBa_K1399007
1
BBa_K1399007
GFP (mut3b) with SsrA-DAS+2 degradation tag
2014-09-18T11:00:00Z
2015-05-08T01:10:16Z
GFP comes from part BBa_E0040, the tag sequence was obtained from part BBa_M0051.
GFP (mut3b) (see part BBa_E0040) with added engineered NYADAS-ssrA degradation tag (see part BBa_M0050). The tag increases GFP turn-over rate, thus providing better temporal resolution of green fluorescence. In the same time, maximal fluorescence amplitudes will be lower as newly formed protein is degraded as soon as it is formed.
SsrA tags encode peptide sequence that and is recognized by ClpA and ClpX unfoldases and ClpX mediator SspB.[1] ClpA and ClpX then form a proteosome-like complex with ClpP protease and the protein is degraded.[1] The final three residues of the tag determines the strength of interaction with ClpX and thus the final protein degradation rate.[2] The NYADAS tag encodes peptide sequence AANDENYNYDAS is reported to have low affinity to ClpX thus its mediated degradation very much depends on the concentration of SspB (ClpX mediator).[1] The two additional residues ???NY??? extends tag between SspB and ClpX binding site, thus preventing clash when both these protein are bound to tag.[3] However, be aware that exact protein degradation rate is influenced by multiple other factors: ClpXP and ClpAP protease concentrations, protein stability, Km of binding to the protease, temperature [4].
===References===
[1] Flynn, J. M. et al. Overlapping recognition determinants within the ssrA degradation tag allow modulation of proteolysis. Proc. Natl. Acad. Sci. U. S. A. 98, 10584???9 (2001).
[2] Andersen, J. B. et al. New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. Appl. Environ. Microbiol. 64, 2240???6 (1998).
[3] McGinness, K. E., Baker, T. a & Sauer, R. T. Engineering controllable protein degradation. Mol. Cell 22, 701???7 (2006).
[4] Purcell, O., Grierson, C. S., Bernardo, M. Di & Savery, N. J. Temperature dependence of ssrA-tag mediated protein degradation. J. Biol. Eng. 6, 10 (2012).
false
false
_1777_
0
22477
9
In stock
true
The tag was attached to GFP using PCR and MABEL (mutagenesis with blunt-end ligation), thus avoiding introduction of additional residues and restriction site. Different parts of the tag are recognized by different proteins, for example, the final 3 residues (DAS in this case) are recognized by ClpX, whereas first 4 residues of the tag are required for efficient SspB binding.[1] Thus modifications of these critical residues alter the efficacy with what different proteases bind to it.
false
Anna Stikane
annotation2383924
1
GFP (mut3b)
range2383924
1
4
714
annotation2383923
1
start
range2383923
1
1
3
annotation2383926
1
stop
range2383926
1
754
756
annotation2383925
1
NYADAS-ssrA
range2383925
1
715
753
annotation2383927
1
stop
range2383927
1
757
759
BBa_K1462950
1
BBa_K1462950
CTS1-1-+GFP
2014-10-14T11:00:00Z
2015-05-08T01:10:36Z
The targetting sequence derived from yeast can be located at cytoderm.
A COOH-terminal 12-amino acid serve as a typical cleavable signal sequence
false
false
_1841_
0
21372
9
Not in stock
false
We add a GFP to make the location visible.
false
LinZhou Li,YaRan Zhao
component2419960
1
BBa_K1462930
component2419966
1
BBa_K1399007
annotation2419960
1
BBa_K1462930
range2419960
1
1
60
annotation2419966
1
BBa_K1399007
range2419966
1
67
825
BBa_K801012_sequence
1
agctttggacttcttcgccagaggtttggtcaagtctccaatcaaggttgtcggcttgtctaccttgccagaaatttacgaaaagatggaaaagggtcaaatcgttggtagatacgttgttgacacttctaaataagcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccggccggtcgaaattcccctaccctatg
BBa_K1462930_sequence
1
atgtcactcctttacatcattcttctattcacacaattcttactactgccaaccgatgcc
BBa_K530008_sequence
1
acagtttattcctggcatccactaaatataatggagcccgctttttaagctggcatccagaaaaaaaaagaatcccagcaccaaaatattgttttcttcaccaaccatcagttcataggtccattctcttagcgcaactacagagaacaggggcacaaacaggcaaaaaacgggcacaacctcaatggagtgatgcaacctgcctggagtaaatgatgacacaaggcaattgacccacgcatgtatctatctcattttcttacaccttctattaccttctgctctctctgatttggaaaaagctgaaaaaaaaggttgaaaccagttccctgaaattattcccctacttgactaataagtatataaagacggtaggtattgattgtaattctgtaaatctatttcttaaacttcttaaattctacttttatagttagtcttttttttagttttaaaacaccaagaacttagtttcgaataaacacacataaacaaacaaa
BBa_K1462970_sequence
1
acagtttattcctggcatccactaaatataatggagcccgctttttaagctggcatccagaaaaaaaaagaatcccagcaccaaaatattgttttcttcaccaaccatcagttcataggtccattctcttagcgcaactacagagaacaggggcacaaacaggcaaaaaacgggcacaacctcaatggagtgatgcaacctgcctggagtaaatgatgacacaaggcaattgacccacgcatgtatctatctcattttcttacaccttctattaccttctgctctctctgatttggaaaaagctgaaaaaaaaggttgaaaccagttccctgaaattattcccctacttgactaataagtatataaagacggtaggtattgattgtaattctgtaaatctatttcttaaacttcttaaattctacttttatagttagtcttttttttagttttaaaacaccaagaacttagtttcgaataaacacacataaacaaacaaatactagatgtcactcctttacatcattcttctattcacacaattcttactactgccaaccgatgcctactagatgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaagctgcaaacgacgaaaactacaactacgctgacgcttcttaataatactagagagctttggacttcttcgccagaggtttggtcaagtctccaatcaaggttgtcggcttgtctaccttgccagaaatttacgaaaagatggaaaagggtcaaatcgttggtagatacgttgttgacacttctaaataagcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccggccggtcgaaattcccctaccctatg
BBa_K1399007_sequence
1
atgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaagctgcaaacgacgaaaactacaactacgctgacgcttcttaataa
BBa_K1462950_sequence
1
atgtcactcctttacatcattcttctattcacacaattcttactactgccaaccgatgcctactagatgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaagctgcaaacgacgaaaactacaactacgctgacgcttcttaataa
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
Chris J. Myers
James Alastair McLaughlin
2017-03-06T15:00:00.000Z