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_K1362438
1
Gly3
GGG linker
2014-10-07T11:00:00Z
2015-05-08T01:10:05Z
Triglycine linker
false
false
_1738_
0
22920
9
Not in stock
false
false
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Büscher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schäfer, Carolin Schmelas, Silvan Schmitz, Max Waldha
BBa_K1362054
1
BBa_K1362054
SsrA degradation tag as RFC[105] C-terminal insert
2014-10-07T11:00:00Z
2015-05-08T01:10:04Z
false
false
_1738_
0
22920
9
In stock
false
false
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena B??scher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Sch??fer, Carolin Schmelas, Silvan Schmitz, Max Waldha
component2409703
1
BBa_K1362426
component2409701
1
BBa_K1362421
component2409694
1
BBa_K1362419
component2409699
1
BBa_M0050
component2409693
1
BBa_K1362425
component2409697
1
BBa_K1362438
component2409702
1
BBa_K1362423
component2409696
1
BBa_K1362433
annotation2409701
1
BBa_K1362421
range2409701
1
62
65
annotation2409702
1
BBa_K1362423
range2409702
1
66
72
annotation2409693
1
BBa_K1362425
range2409693
1
1
6
annotation2409696
1
BBa_K1362433
range2409696
1
11
19
annotation2409699
1
BBa_M0050
range2409699
1
29
61
annotation2409697
1
BBa_K1362438
range2409697
1
20
28
annotation2409703
1
BBa_K1362426
range2409703
1
73
76
annotation2409694
1
BBa_K1362419
range2409694
1
7
10
BBa_K1362433
1
CWE
CWE (extein)
2014-10-07T11:00:00Z
2015-05-08T01:10:05Z
???
???
false
false
_1738_
0
22920
9
Not in stock
false
???
false
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Büscher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schäfer, Carolin Schmelas, Silvan Schmitz, Max Waldha
annotation2407307
1
CWE
range2407307
1
1
9
BBa_K1362421
1
RFC105 Z
C-terminal stop overhang TAAT=STOP+1/3Stop RFC[105] overhang Z
2014-10-06T11:00:00Z
2015-05-08T01:10:05Z
This standard overhang part was developed as part of the iGEM team Heidelberg 2014's Intein Toolbox [[#References|[1]]]. All standard sequences can be reviewed in RFC[???] [[#References|[2]]].
This is a standard overhang sequence for in-frame cloning of Proteins of Interest in front or behind an Intein. A detailed cloning strategy is found on the iGEM team Heidelberg 2014's [http://2014.igem.org/Team:Heidelberg/parts wiki page] as well as in RFC[???]. Specifically, this part contains the overhang F used to insert the last part of an intein-fused protein directly in front of an RFC[10] double stop-codon. It lies within the first for bases of the stopstop sequence.
false
false
_1738_
0
12377
9
Not in stock
false
This part is only the sequence of a standard overhang. It will not be sent in as physical DNA and was merely created to easily compose new parts in the RFC[???] standard. In the actual cloning process this sequence was and is recommended to be inserted with the according PCR primers.
false
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena B??scher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Sch??fer, Carolin Schmelas, Silvan Schmitz, Max Waldha
annotation2402257
1
stop
range2402257
1
1
4
BBa_K1362426
1
spacer
Random sequence
2014-10-07T11:00:00Z
2015-05-08T01:10:05Z
It was suddenly there.
This sequence is of no use.
false
false
_1738_
0
22830
9
Not in stock
false
No consideration was performed by the designer.
false
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Büscher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schäfer, Carolin Schmelas, Silvan Schmitz, Max Waldha
BBa_K1362425
1
BsaI ->
BsaI restriction site for RFC[???] cloning (in part in prefix)
2014-10-07T11:00:00Z
2015-05-08T01:10:05Z
This Sequence starts with a part of the BsaI recognition site. The missing G of the recognition site can be found in the upstream BB prefix. It contains a spacer nucleotide so that BsaI will cut the top strand directly downstream and the bottom strand 4 nucleotides downstream.
false
false
_1738_
0
22830
9
Not in stock
false
false
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Büscher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schäfer, Carolin Schmelas, Silvan Schmitz, Max Waldha
annotation2402282
1
RFC[???] standard overhang A
range2402282
1
5
5
annotation2400588
1
BsaI site
range2400588
1
1
5
BBa_K1362419
1
RFC105 CC
C-terminal splicing overhang CAAC=1/3?+Asn RFC[105] standard overhang CC
2014-10-06T11:00:00Z
2015-05-08T01:10:05Z
This standard overhang part was developed as part of the iGEM team Heidelberg 2014's Intein Toolbox [[#References|[1]]]. All standard sequences can be reviewed in RFC[???] [[#References|[2]]].
This is a standard overhang sequence for in-frame cloning of Proteins of Interest in front or behind an Intein. A detailed cloning strategy is found on the iGEM team Heidelberg 2014's [http://2014.igem.org/Team:Heidelberg/parts wiki page] as well as in RFC[???]. Specifically, this part contains the overhang E used to insert a protein in front of an N-Intein. It lies within the four bases formed by the cystein and the first base of the subsequent Leucine/Isoleucine or similar of the N-terminal splicing site.
false
false
_1738_
0
12377
9
Not in stock
false
This part is only the sequence of a standard overhang. It will not be sent in as physical DNA and was merely created to easily compose new parts in the RFC[???] standard. In the actual cloning process this sequence was and is recommended to be inserted with the according PCR primers.
false
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena B??scher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Sch??fer, Carolin Schmelas, Silvan Schmitz, Max Waldha
BBa_K1362423
1
<- BsaI
BsaI reverse restriction site for RFC[105] cloning
2014-10-06T11:00:00Z
2015-05-08T01:10:05Z
This standard restriction site sequence is used as part of the iGEM team Heidelberg 2014's Intein Toolbox [[#References|[1]]]. All associated standard sequences can be reviewed in RFC[???]
This is the reverse complement of a BsaI restriction site headed by an Adenine as a spacer-base to separate the recognition sequence from the outward-lying cutting sequence. It was used by us for scarless golden-gate cloning to fuse inteins to other proteins and thereby implement a variety of possible port-translational modifications.
false
false
_1738_
0
12377
9
Not in stock
false
This part is only the sequence of a restriction site. It will not be sent in as physical DNA and was merely created to easily compose new parts in the RFC[???] standard. In the actual cloning process this sequence was and is recommended to be inserted with the according PCR primers.
false
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena B??scher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Sch??fer, Carolin Schmelas, Silvan Schmitz, Max Waldha
BBa_K1362425_sequence
1
gtctcc
BBa_K1362426_sequence
1
tgtc
BBa_K1362419_sequence
1
caac
BBa_K1362423_sequence
1
agagacc
BBa_K1362433_sequence
1
tgctgggaa
BBa_K1362438_sequence
1
ggaggaggt
BBa_K1362054_sequence
1
gtctcccaactgctgggaaggaggaggtgctgctaacgacgaaaactacgctctggctgcttaatagagacctgtc
BBa_K1362421_sequence
1
taat
BBa_M0050_sequence
1
gctgctaacgacgaaaactacgctctggctgct
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