BBa_M0050
1
LAA
AANDENYALAA. (Very fast) SsrA degradation tag.
2007-12-05T12:00:00Z
2015-05-08T01:13:51Z
C-terminal degradation tags are commonly found in high turnover proteins in Escherichia coli.
This sequence codes for the amino acid sequence AANDENYALAA, which when fused to the C-terminal of proteins, will make the protein susceptible to very fast degradation through SspB-mediated binding to the ClpX protease.
The following rates of degradation of this tag are pulled from the corresponding references below:
~5 Vmax/ [Clpx6] min-1 from (1)
~0.5%/min on log scale from (2)
~1 min half life from (3)
See the following references for further information on degradation rates and mechanisms of this tag:
(1)McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701.
(2)Flynn et al 2003. Mol. Cell. 11: 671.
Flynn et al. 2001. PNAS 98(19): 10584.
Anderson et al 1998. App. Env. Microbiol. 64(6):2240.
false
false
_11_
0
2398
11
Not in stock
false
C-terminal tag.
Degradation rate is very fast. Deviations from this sequence in key amino acids will lower degradation rates (see Parts BBa_M0051, BBa_M0052, BBa_M0053).
Three C-terminal aa's (LAA in this case) are necessary and sufficient for ClpX binding and degradation. Upstream aa sequence serves as a binding site for SspB, which guides rapid binding to ClpX.
false
Felix Moser
annotation1958880
1
WT SsrA tag AANDENYALAA
range1958880
1
1
33
BBa_B0015
1
BBa_B0015
double terminator (B0010-B0012)
2003-07-16T11:00:00Z
2015-08-31T04:07:20Z
Released HQ 2013
Double terminator consisting of BBa_B0010 and BBa_B0012
false
true
_1_
0
24
7
In stock
false
true
Reshma Shetty
component1916610
1
BBa_B0010
component1916612
1
BBa_B0012
annotation1916610
1
BBa_B0010
range1916610
1
1
80
annotation1916612
1
BBa_B0012
range1916612
1
89
129
BBa_K743018
1
sfGFP.degr
sfGFP.degradation.tag under Pta promoter in psb1C3_IntKR plasmid
2012-09-22T11:00:00Z
2015-05-08T01:13:09Z
false
false
_994_
0
11584
9
It's complicated
false
false
component2194674
1
BBa_K743001
component2194654
1
BBa_K743003
component2194658
1
BBa_I746916
component2194652
1
BBa_K743000
component2194660
1
BBa_M0050
component2194672
1
BBa_S05073
annotation2194660
1
BBa_M0050
range2194660
1
1415
1447
annotation2194658
1
BBa_I746916
range2194658
1
687
1406
annotation2194654
1
BBa_K743003
range2194654
1
531
680
annotation2194674
1
BBa_K743001
range2194674
1
2568
3254
annotation2194652
1
BBa_K743000
range2194652
1
1
522
annotation2194672
1
BBa_S05073
range2194672
1
1456
2559
BBa_P1007
1
KanR
kanamycin resistance cassette
2006-02-03T12:00:00Z
2015-05-08T01:14:11Z
This contains KanR, a terminator and then AmpR and a terminator.
false
false
_41_6_
0
126
45
Not in stock
false
false
Reshma Shetty
annotation1919399
1
kanamycin resistance
range1919399
1
1
819
annotation1919400
1
G to C mutation to eliminate XhoI site
range1919400
1
787
787
annotation1919401
1
repeat region
range1919401
1
924
967
BBa_K743001
1
RS2
Synechocystis PCC6803 neutral recombination site.
2012-09-19T11:00:00Z
2015-05-08T01:13:09Z
PCR amplified from Synechocystis PCC6803 chromosome, orf slr0337
When placed between this part and BBa_K743000, any DNA sequence will be inserted into Synechocystis PCC6803 chromosome trough homologous recombination
false
false
_994_
0
11584
9
It's complicated
false
According to literature, in Synechocystis the recombination sites must be at least 450 bp long to allow a rasonably high transformation efficiency, being approximately 1200 bp the optimum size.
false
annotation2188524
1
RS2
range2188524
1
1
687
BBa_I746916
1
BBa_I746916
superfolder GFP coding sequence
2008-09-29T11:00:00Z
2015-08-31T04:08:05Z
Superfolder GFP was originally described by: Pedelacq et al (2006): "Engineering and characterization of a superfolder green fluorescent protein", Nature Biotech 24 (1) January 2006
This version was synthesised de novo (by Geneart).
This is the coding sequence of superfolder GFP (Pedelacq et al (2006): "Engineering and characterization of a superfolder green fluorescent protein", Nature Biotech 24 (1) January 2006).
It carries the following amino acid changes with respect to mut3 GFP (E0040), the currently most commonly used GFP in the registry:
S30R, Y39N, F64L, G65T, F99S, N105T, Y145F, M153T, V163A, I171V, A206V
Its in-vivo properties are considerably improved with respect to mut3 - it develops fluorescence about 3fold faster than mut3 GFP and reaches 4fold higher absolute fluorescence levels. Fluorescenct colonies can be identified with the naked eye even without UV or blue light illumination (that is to say the amount of blue light in normal daylight or lablight is sufficient).
Additionally it is more stable in vitro and refolds faster after in vitro denaturation with respect to mut3 GFP.
Note:
Superfolder GFP is available in constructs driven by the pBAD and T7 promoters: part numbers I746908 and I746909 respectively. Additionally 6-his tagged versions for protein purification exist: I746914 (pBAD driven) and I746915 (T7 driven).
false
false
_116_
0
2122
9
It's complicated
false
Codon optimisation before de novo synthesis was carried out for both, E.coli and Bacillus subtilis.
false
Stefan Milde
annotation1977535
1
stop
range1977535
1
715
720
annotation1977533
1
start
range1977533
1
1
3
annotation1977534
1
superfolder GFP coding region
range1977534
1
1
720
BBa_K743003
1
BBa_K743003
Pta promoter and native rbs from Synechocystis PCC6803
2012-09-19T11:00:00Z
2015-05-08T01:13:09Z
This part was PCR amplified from Synechocystis chromosome, orf slr1739.
This is the 150bp sequence upstream the Transaldolase gene start codon, it contains putative rbs and promoter regions.
Transaldolase is an enzyme involved in the pentose phospate cycle and its mRNA levels have shown to oscillate in a circadian manner, peaking at hour 14.
false
false
_994_
0
11584
9
Not in stock
false
It was assumed that the 150bp sequence upstream from the start codon includes the promoter and the ribosome binding site
false
Juan Sim??n ??lamos
annotation2188682
1
Pta
range2188682
1
1
150
BBa_K743000
1
RS1
Synechocystis PCC6803 neutral recombination site.
2012-09-19T11:00:00Z
2015-05-08T01:13:09Z
PCR amplified from synechocystis PCC6803 chromosome.
Any DNA sequence flacked by this part and BBa_K743001 will be incorporated into Synechocistys PCC6803 chromosome through a neutral double homologous recombination.
false
false
_994_
0
11584
9
In stock
false
According to literature, in Synechocystis the recombination sites must be at least 450bp long to allow a rasonably high transformation efficiency, being 700bp the optimum size.
false
annotation2188518
1
RS1 recombination site
range2188518
1
1
522
BBa_S05073
1
BBa_S05073
B0015:P1007
2012-09-19T11:00:00Z
2015-05-08T01:14:49Z
false
false
_9_
0
11584
9
Not in stock
false
false
Juan Sim??n ??lamos
component2188869
1
BBa_B0015
component2188873
1
BBa_P1007
annotation2188869
1
BBa_B0015
range2188869
1
1
129
annotation2188873
1
BBa_P1007
range2188873
1
138
1104
BBa_B0010
1
BBa_B0010
T1 from E. coli rrnB
2003-11-19T12:00:00Z
2015-08-31T04:07:20Z
Transcriptional terminator consisting of a 64 bp stem-loop.
false
false
_1_
0
24
7
In stock
false
true
Randy Rettberg
annotation7018
1
BBa_B0010
range7018
1
1
80
annotation4184
1
stem_loop
range4184
1
12
55
BBa_B0012
1
BBa_B0012
TE from coliphageT7
2003-01-31T12:00:00Z
2015-08-31T04:07:20Z
Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>.
Released HQ 2013
Transcription terminator for the <i>E.coli</i> RNA polymerase.
false
false
_1_
0
24
7
In stock
false
<P> <P>Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG).<P> Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator.
false
Reshma Shetty
annotation1690
1
polya
range1690
1
28
41
annotation7020
1
BBa_B0012
range7020
1
1
41
annotation1687
1
stop
range1687
1
34
34
annotation1686
1
T7 TE
range1686
1
8
27
BBa_B0010_sequence
1
ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctc
BBa_K743001_sequence
1
agtgtaggcggtaaagtctatgggaacttccagtttaagggctgtcccgttaaatatggcttcgagtttttgaagcataattttctaacgtctgtggatttttttaagcatagcttgaccttctgttggtcgaattctagggattttcatccatattcggttcatagggcagctcaatggtgaacattgctcccccagtttcatggttagctgcggttaaatttcccctgtgggccaggacaatttccttggcgatcgccaaacccaggccactgccatggcgttgactatcgggatgtaggcgagcacgggaactatcaccccgataaagtcgctcaaaaatgtagggtaaatcccttggttgaaatcctaggccttgatccctaataatcatggtgatcccttctgtgccttgatgaccctgcacaaaaatagtcccttccggtgggctatatttcagcgtattgtccaaaatattcatcaatacctgcatcagacgatcgccatccccctctaggttcaacttggttggcccttggtaatccagggtgatattttttaccacggcgatcggggttaatcgctcccaggtaatggcgagtaaatggcgtaaattaatcggttctggctcgaggtaaagattgggattagcggttatttgggttaaatggagccaactttccacca
BBa_K743018_sequence
1
tcccagcctctcaaccaccgatattgggcccctagctacccctgatattttggcggacattgttgcgcaagtagaacaaaccattgctgctggagcccactgtcgttgtggcggccaagccctagaccaaccgggaaattactatcctcccaccctgctcaccgacgttccccccaacgcccctacctaccgtcaggaattttttggccccgtggccctcggatttactgtcgataatttagaggaggcgatcgccttggccaatgacattccctttgggttgggggccagtgcatggacaactaacccggaaaatcaacaaaaactaatccggggcatagaagccggggctgtattcattaacggtatgactaaatctgacccccgcattccctttggaggcatcaagcgctctggctttgggcgggaacttggccgcatgggcattttagaatttgtcaatgctaaaaccgtttggattgcttagtccgccaccattttaaaattatcgctcttctattctactagagaacgcatcggcgttccctagcttaaattatcttgatgtccaaagaaggctcgctttcgggaatcaccattaccattggagggaaagacgttgaacttgtttaatcgccattatgggtaagaatctcctcgaacaactgcgccaatttacatactagatgcgtaaaggcgaagagctgttcactggtgtcgtccctattctggtggaactggatggtgatgtcaacggtcataagttttccgtgcgtggcgagggtgaaggtgacgcaactaatggtaaactgacgctgaagttcatctgtactactggtaaactgccggtaccttggccgactctggtaacgacgctgacttatggtgttcagtgctttgctcgttatccggaccatatgaagcagcatgacttcttcaagtccgccatgccggaaggctatgtgcaggaacgcacgatttcctttaaggatgacggcacgtacaaaacgcgtgcggaagtgaaatttgaaggcgataccctggtaaaccgcattgagctgaaaggcattgactttaaagaagacggcaatatcctgggccataagctggaatacaattttaacagccacaatgtttacatcaccgccgataaacaaaaaaatggcattaaagcgaattttaaaattcgccacaacgtggaggatggcagcgtgcagctggctgatcactaccagcaaaacactccaatcggtgatggtcctgttctgctgccagacaatcactatctgagcacgcaaagcgttctgtctaaagatccgaacgagaaacgcgatcatatggttctgctggagttcgtaaccgcagcgggcatcacgcatggtatggatgaactgtacaaatgatgatactagaggctgctaacgacgaaaactacgctctggctgcttactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagttattagaaaaactcatcgagcatcaaatgaaactgcaatttattcatatcaggattatcaataccatatttttgaaaaagccgtttctgtaatgaaggagaaaactcaccgaggcagttccaaagaatggcaaggtcctggtaacggtctgcgattccgacccgtccaacatcaatacaacctattaatttcccctcgtcaaaaataaggttatcaagtgagaaatcaccatgagtgacgactgaatccggtgagaatggcaagagcttgtgcatttctttccagacttgttcaacaggccagccattacgctcgtcatcaaaatcactcgcatcaaccaaaccgttattcatgcgtgattgcgcctgagcaagacgaaatacacgatcgctgttaaaaggacaattacaaacaggaatcgaatgtaaccggcgcaggaacacggccagcgcatcaacaatattttcacctgaatcaggatattcttctaatacctggaaggctgttttcccaggaatcgcggtggtgagtaaccacgcatcatcaggagtacggataaaatgcttgatggtcgggagaggcataaactccgtcagccagttgagacggaccatctcatctgtaacatcattggcaacgctacctttgccatgtttcagaaacaactctggcgcatcgggcttcccatacaagcgatagattgtcgcacctgattgcccgacattatcgcgagcccatttatacccatataaatcagcgtccatgttggagtttaagcgcggacgggagcaagacgtttcccgttgaatatggctcataacaccccttgtattactgtttatgtaagcagacagttttattgttcatgatgatatatttttatcttgtgcaatgtaacatcagagattttgagacacaacgtggctttgttgaataaatcgaacttttgctgagttgaaggatcagtactagagagtgtaggcggtaaagtctatgggaacttccagtttaagggctgtcccgttaaatatggcttcgagtttttgaagcataattttctaacgtctgtggatttttttaagcatagcttgaccttctgttggtcgaattctagggattttcatccatattcggttcatagggcagctcaatggtgaacattgctcccccagtttcatggttagctgcggttaaatttcccctgtgggccaggacaatttccttggcgatcgccaaacccaggccactgccatggcgttgactatcgggatgtaggcgagcacgggaactatcaccccgataaagtcgctcaaaaatgtagggtaaatcccttggttgaaatcctaggccttgatccctaataatcatggtgatcccttctgtgccttgatgaccctgcacaaaaatagtcccttccggtgggctatatttcagcgtattgtccaaaatattcatcaatacctgcatcagacgatcgccatccccctctaggttcaacttggttggcccttggtaatccagggtgatattttttaccacggcgatcggggttaatcgctcccaggtaatggcgagtaaatggcgtaaattaatcggttctggctcgaggtaaagattgggattagcggttatttgggttaaatggagccaactttccacca
BBa_K743000_sequence
1
tcccagcctctcaaccaccgatattgggcccctagctacccctgatattttggcggacattgttgcgcaagtagaacaaaccattgctgctggagcccactgtcgttgtggcggccaagccctagaccaaccgggaaattactatcctcccaccctgctcaccgacgttccccccaacgcccctacctaccgtcaggaattttttggccccgtggccctcggatttactgtcgataatttagaggaggcgatcgccttggccaatgacattccctttgggttgggggccagtgcatggacaactaacccggaaaatcaacaaaaactaatccggggcatagaagccggggctgtattcattaacggtatgactaaatctgacccccgcattccctttggaggcatcaagcgctctggctttgggcgggaacttggccgcatgggcattttagaatttgtcaatgctaaaaccgtttggattgcttagtccgccaccattttaaaattatcgctcttctattc
BBa_I746916_sequence
1
atgcgtaaaggcgaagagctgttcactggtgtcgtccctattctggtggaactggatggtgatgtcaacggtcataagttttccgtgcgtggcgagggtgaaggtgacgcaactaatggtaaactgacgctgaagttcatctgtactactggtaaactgccggtaccttggccgactctggtaacgacgctgacttatggtgttcagtgctttgctcgttatccggaccatatgaagcagcatgacttcttcaagtccgccatgccggaaggctatgtgcaggaacgcacgatttcctttaaggatgacggcacgtacaaaacgcgtgcggaagtgaaatttgaaggcgataccctggtaaaccgcattgagctgaaaggcattgactttaaagaagacggcaatatcctgggccataagctggaatacaattttaacagccacaatgtttacatcaccgccgataaacaaaaaaatggcattaaagcgaattttaaaattcgccacaacgtggaggatggcagcgtgcagctggctgatcactaccagcaaaacactccaatcggtgatggtcctgttctgctgccagacaatcactatctgagcacgcaaagcgttctgtctaaagatccgaacgagaaacgcgatcatatggttctgctggagttcgtaaccgcagcgggcatcacgcatggtatggatgaactgtacaaatgatga
BBa_K743003_sequence
1
aacgcatcggcgttccctagcttaaattatcttgatgtccaaagaaggctcgctttcgggaatcaccattaccattggagggaaagacgttgaacttgtttaatcgccattatgggtaagaatctcctcgaacaactgcgccaatttaca
BBa_B0012_sequence
1
tcacactggctcaccttcgggtgggcctttctgcgtttata
BBa_M0050_sequence
1
gctgctaacgacgaaaactacgctctggctgct
BBa_B0015_sequence
1
ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata
BBa_P1007_sequence
1
ttattagaaaaactcatcgagcatcaaatgaaactgcaatttattcatatcaggattatcaataccatatttttgaaaaagccgtttctgtaatgaaggagaaaactcaccgaggcagttccaaagaatggcaaggtcctggtaacggtctgcgattccgacccgtccaacatcaatacaacctattaatttcccctcgtcaaaaataaggttatcaagtgagaaatcaccatgagtgacgactgaatccggtgagaatggcaagagcttgtgcatttctttccagacttgttcaacaggccagccattacgctcgtcatcaaaatcactcgcatcaaccaaaccgttattcatgcgtgattgcgcctgagcaagacgaaatacacgatcgctgttaaaaggacaattacaaacaggaatcgaatgtaaccggcgcaggaacacggccagcgcatcaacaatattttcacctgaatcaggatattcttctaatacctggaaggctgttttcccaggaatcgcggtggtgagtaaccacgcatcatcaggagtacggataaaatgcttgatggtcgggagaggcataaactccgtcagccagttgagacggaccatctcatctgtaacatcattggcaacgctacctttgccatgtttcagaaacaactctggcgcatcgggcttcccatacaagcgatagattgtcgcacctgattgcccgacattatcgcgagcccatttatacccatataaatcagcgtccatgttggagtttaagcgcggacgggagcaagacgtttcccgttgaatatggctcataacaccccttgtattactgtttatgtaagcagacagttttattgttcatgatgatatatttttatcttgtgcaatgtaacatcagagattttgagacacaacgtggctttgttgaataaatcgaacttttgctgagttgaaggatcag
BBa_S05073_sequence
1
ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagttattagaaaaactcatcgagcatcaaatgaaactgcaatttattcatatcaggattatcaataccatatttttgaaaaagccgtttctgtaatgaaggagaaaactcaccgaggcagttccaaagaatggcaaggtcctggtaacggtctgcgattccgacccgtccaacatcaatacaacctattaatttcccctcgtcaaaaataaggttatcaagtgagaaatcaccatgagtgacgactgaatccggtgagaatggcaagagcttgtgcatttctttccagacttgttcaacaggccagccattacgctcgtcatcaaaatcactcgcatcaaccaaaccgttattcatgcgtgattgcgcctgagcaagacgaaatacacgatcgctgttaaaaggacaattacaaacaggaatcgaatgtaaccggcgcaggaacacggccagcgcatcaacaatattttcacctgaatcaggatattcttctaatacctggaaggctgttttcccaggaatcgcggtggtgagtaaccacgcatcatcaggagtacggataaaatgcttgatggtcgggagaggcataaactccgtcagccagttgagacggaccatctcatctgtaacatcattggcaacgctacctttgccatgtttcagaaacaactctggcgcatcgggcttcccatacaagcgatagattgtcgcacctgattgcccgacattatcgcgagcccatttatacccatataaatcagcgtccatgttggagtttaagcgcggacgggagcaagacgtttcccgttgaatatggctcataacaccccttgtattactgtttatgtaagcagacagttttattgttcatgatgatatatttttatcttgtgcaatgtaacatcagagattttgagacacaacgtggctttgttgaataaatcgaacttttgctgagttgaaggatcag
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