BBa_J18912
1
T7start
T7 promoter + RBS + START
2010-01-26T12:00:00Z
2015-08-31T04:08:36Z
pET3a expression vector
Strong T7 promoter and RBS including start codon for protein expression in E. coli or in-vitro.
Part is provided in RFC 25 assembly format.
false
false
_165_
0
2175
165
It's complicated
false
* sequence is taken from the commonly used pET3a expression vector
* deleted XbaI site between prom. and RBS
* T7 promoter sequence (1-19) equals the one in BBa_I712074
* Conservative choice of part boundaries and modifications -- the part's size could probably be reduced.
false
Raik Gruenberg
annotation2065586
1
start
range2065586
1
81
83
BBa_J18909
1
HisTag
His tag (6xHis; E. coli - optimized)
2010-01-15T12:00:00Z
2015-08-31T04:08:35Z
Synthetic DNA
Hexahistidine affinity tag.
Binds zinc, nickel, or cobalt affinity resins.
=Notes=
This part is identical to BBa_I757013 (which does not seem to be available though).
=References=
Wikipedia article: [http://en.wikipedia.org/wiki/Polyhistidine-tag]
false
false
_165_
0
2175
165
It's complicated
false
Simple repetition of the His codon most frequent in E. coli -- a more balanced codon-choice may be better.
false
Raik Gruenberg
annotation2065463
1
PolyHis
range2065463
1
1
18
BBa_J18925
1
FKBP12
FKBP12
2010-01-26T12:00:00Z
2015-08-31T04:08:36Z
literature
The immunophilin protein FKBP12 (FK506-binding protein) is the target of the natural compounds FK506 and rapamycin. The FKBP/rapamycin or FKBP/FK506 complex then binds to several other proteins and suppresses immune reactions. Drug screening and design has turned up a selection of other small molecules that bind with high specificity to FKBP12 itself or to mutants of FKBP12. A second human protein, FRB is among the most prominent binding targets of the FKBP/rapamycin complex (using the "other face" of the rapamycin molecule). Rapamycin can herefore induce the hetero-dimerization of FKBP12 and FRB.
== See also ==
* structure of FRB / FKBP / Rapamycin complex: PDB 1nsg
(CRG-internal ID: rg0073)
false
false
_165_
0
2175
165
It's complicated
false
* gene synthesis
* codon optimized for E. coli
false
Raik Gruenberg
BBa_J18939
1
BBa_J18939
FKBP ~ bla-frag2
2010-01-26T12:00:00Z
2015-08-31T04:08:36Z
synthetic
P01 expression cassette
from Gruenberg et al. 2010 series of interaction reporter proteins. All proteins share the same architecture:
* T7Start-domain1-5xGS-domain2-preSCsite-HisTag-T7Stop
Only Domains1 and 2 vary and are taken from a list of protein interaction input and output (readout) parts.
===Expression===
Proteins were expressed in E. coli BL21 using T7 Polymerase and were purified by His-tag affinity.
===Reference===
* Gruenberg, Ferrar, van der Sloot, Serrano (2010): Building Blocks for Protein Interaction Devices
false
false
_165_
0
2175
165
It's complicated
false
Constructs were assembled according to R.F.C. 25.
All parts were codon optimized for E. coli.
false
Raik Gruenberg
component2066518
1
BBa_J18922
component2066527
1
BBa_J18919
component2066524
1
BBa_I757012
component2066514
1
BBa_B0105
component2066515
1
BBa_J18925
component2066533
1
BBa_B0105
component2066517
1
BBa_B0105
component2066535
1
BBa_J18913
component2066512
1
BBa_J18912
component2066529
1
BBa_B0105
component2066531
1
BBa_J18909
component2066526
1
BBa_B0105
component2066520
1
BBa_B0105
annotation2066524
1
BBa_I757012
range2066524
1
453
740
annotation2066514
1
BBa_B0105
range2066514
1
84
89
annotation2066512
1
BBa_J18912
range2066512
1
1
83
annotation2066527
1
BBa_J18919
range2066527
1
747
770
annotation2066529
1
BBa_B0105
range2066529
1
771
776
annotation2066520
1
BBa_B0105
range2066520
1
447
452
annotation2066515
1
BBa_J18925
range2066515
1
90
410
annotation2066518
1
BBa_J18922
range2066518
1
417
446
annotation2066535
1
BBa_J18913
range2066535
1
801
935
annotation2066526
1
BBa_B0105
range2066526
1
741
746
annotation2066517
1
BBa_B0105
range2066517
1
411
416
annotation2066531
1
BBa_J18909
range2066531
1
777
794
annotation2066533
1
BBa_B0105
range2066533
1
795
800
BBa_J18913
1
T7stop
T7 terminator (incl. STOP)
2010-01-26T12:00:00Z
2015-08-31T04:08:36Z
pET3a expression plasmid
T7 terminator for protein expression in E. coli or in vitro.
Part is provided in R.F.C. 25 assembly format.
false
false
_165_
0
2175
165
It's complicated
false
* taken from pET3a expression plasmid
* added two STOP to 5'
* conservative choice of part boundary, could probably be shortened
false
Raik Gruenberg
annotation2065587
1
stop
range2065587
1
1
6
BBa_J18922
1
5xGS
10aa [GS]x linker
2010-01-26T12:00:00Z
2015-08-31T04:08:36Z
gene synthesis
Highly flexible 10 amino acid linker. Translates to [GS]_5. Codon-optimize for E. coli, yeast, mammalian.
false
true
_165_
0
2175
165
It's complicated
false
* gene synthesis
* codon optimized for E. coli
* experimental distance distributions are described in Neuweiler et al. 2006 papers.
false
Raik Gruenberg
BBa_I757012
1
BBa_I757012
bla_frag2 (aa 200-290) Fusion Part
2007-10-23T11:00:00Z
2015-08-31T04:08:07Z
synthetic DNA by GeneArt
* second part of the split enzyme beta-lactamase
* NgoMIV / AgeI protein fusion part
* iGEM Team Freiburg 2007
false
false
_
0
2006
9
It's complicated
false
between BioBrick 1.0 sites part is flanked 5' by NgoMIV and 3' AgeI(=PinAI) to facilitate in frame cloning for protein fusions
false
Freiburg iGEM team 2007 and Jochen Hecky
annotation1954463
1
NgoMIV
range1954463
1
4
9
annotation1954464
1
AgeI
range1954464
1
280
285
annotation1955422
1
bla frag2 200-290 (numbering according to Ambler)
range1955422
1
13
279
BBa_B0105
1
Scar 25
RFC 25 Scar Sequence
2009-10-14T11:00:00Z
2015-08-31T04:07:21Z
BBF RFC 25
This is the scar produced by assembly using RFC 25.
If you are assembling a composite part using RFC 25, you can insert this part and specify blunt assembly to get the desired sequence.
false
false
_1_
0
25
397
Not in stock
false
Simple DNA sequence
false
Randy Rettberg
annotation2041621
1
Scar 25
range2041621
1
1
6
BBa_J18919
1
preSCsite
preScission protease cleavage site (E. coli-optimized)
2010-01-26T12:00:00Z
2015-08-31T04:08:36Z
gene synthesis
cleavage site for PreScission??? protease.
preScission protease cleavage site
(PreScission??? protease is a genetically engineered form of human rhinovirus 3C protease with a GST fusion, allowing for facile cleavage and purification of GST-tagged proteins along with protease removal after recombinant protein cleavage. Enables low-temperature cleavage of fusion proteins containing the eight-residue recognition sequence.)
cleavage site:
* Leu-Glu-Val-Leu-Phe-Gln↓Gly-Pro
References
==============
Walker et al. (1994): Efficient and rapid affinity purification of proteins using recombinant fusion proteases. PMID: 7764949
false
false
_165_
0
2175
165
It's complicated
false
* gene synthesis
* codon optimized for E. coli
false
Raik Gruenberg
BBa_J18922_sequence
1
ggtagcggcagcggtagcggtagcggcagc
BBa_B0105_sequence
1
accggc
BBa_J18913_sequence
1
tagtgactgctaacaaagcccgaaaggaagctgagttggctgctgccaccgctgagcaataactagcataaccccttggggcctctaaacgggtcttgaggggttttttgctgaaaggaggaactatatccggat
BBa_J18909_sequence
1
catcatcatcatcatcat
BBa_I757012_sequence
1
atggccggcggtactccggcttcccggcaacaattaatggactggatgaaagcggataaagttgcaggaccacttctgcgctcggtgcttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaactgaatcgtcagatcgctgagataggtgcctcactgattaagcattggaccggttaa
BBa_J18912_sequence
1
taatacgactcactatagggagaccacaacggtttccctctagcaataattttgtttaactttaagaaggagatatacatatg
BBa_J18939_sequence
1
taatacgactcactatagggagaccacaacggtttccctctagcaataattttgtttaactttaagaaggagatatacatatgaccggcggtgtgcaggtggaaaccattagcccgggtgatggccgtacctttccgaaacgtggccagacctgcgtggtgcattataccggcatgctggaagatggcaaaaaatttgatagcagccgcgatcgtaacaaaccgttcaaattcatgctgggcaaacaggaagtgattcgcggctgggaggaaggcgtggcgcagatgagcgttggccagcgtgcgaaactgaccatcagcccggattatgcgtatggcgcgaccggccatccgggtattattccgccgcatgcgaccctggtgtttgatgtggaactgctgaaactggaaaccggcggtagcggcagcggtagcggtagcggcagcaccggcatggccggcggtactccggcttcccggcaacaattaatggactggatgaaagcggataaagttgcaggaccacttctgcgctcggtgcttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaactgaatcgtcagatcgctgagataggtgcctcactgattaagcattggaccggttaaaccggcctggaagtgctgtttcagggcccgaccggccatcatcatcatcatcataccggctagtgactgctaacaaagcccgaaaggaagctgagttggctgctgccaccgctgagcaataactagcataaccccttggggcctctaaacgggtcttgaggggttttttgctgaaaggaggaactatatccggat
BBa_J18925_sequence
1
ggtgtgcaggtggaaaccattagcccgggtgatggccgtacctttccgaaacgtggccagacctgcgtggtgcattataccggcatgctggaagatggcaaaaaatttgatagcagccgcgatcgtaacaaaccgttcaaattcatgctgggcaaacaggaagtgattcgcggctgggaggaaggcgtggcgcagatgagcgttggccagcgtgcgaaactgaccatcagcccggattatgcgtatggcgcgaccggccatccgggtattattccgccgcatgcgaccctggtgtttgatgtggaactgctgaaactggaa
BBa_J18919_sequence
1
ctggaagtgctgtttcagggcccg
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