BBa_B0014
1
BBa_B0014
double terminator (B0012-B0011)
2003-07-15T11:00:00Z
2015-08-31T04:07:20Z
Released HQ 2013
Double terminator consisting of BBa_B0012 and BBa_B0011
false
true
_1_
0
24
7
In stock
false
true
Reshma Shetty
component939303
1
BBa_B0012
component939311
1
BBa_B0011
annotation939303
1
BBa_B0012
range939303
1
1
41
annotation939311
1
BBa_B0011
range939311
1
50
95
BBa_B0011
1
BBa_B0011
LuxICDABEG (+/-)
2003-01-31T12:00:00Z
2015-08-31T04:07:20Z
Derived from luxICDABEG operon terminator of Vibrio fischeri <genbank>AF170104</genbank>.
Released HQ 2013
Bidirectional transcriptional terminator consisting of a 22 bp stem-loop.</p>
false
false
_1_
0
24
7
In stock
false
<P> <P>In the naturally-occuring sequence there is a mismatch in the stem of the stem loop. This can be corrected via an A->G mutation (at position 40 -- sequence coordinate/not MFOLD coordinate). The above sequence does not reflect this mutation (but the MFOLD image does). This terminator's location cannot be found using some inverted repeat detectors like PALINDROME because it is too short and contains a mismatch. This one was found with the help of Tom Knight. It lies between two coding regions that point towards eachother.<P>
true
Reshma Shetty
annotation7019
1
BBa_B0011
range7019
1
1
46
annotation1683
1
stem_loop
range1683
1
13
35
BBa_K316043
1
LytC-Flex-
LytC - Flexible Linker - TEV Cleavage Site - AIP - His Tag
2010-10-26T11:00:00Z
2015-05-08T01:11:57Z
source
intro
false
false
_440_
0
7480
9
Not in stock
false
design
false
IC 2010 Team
annotation2109248
1
Flexible Linker
range2109248
1
977
1050
annotation2109245
1
sRBS
range2109245
1
1
15
annotation2109253
1
Stop Codon
range2109253
1
1177
1179
annotation2109250
1
Auto Inducing Peptide
range2109250
1
1105
1155
annotation2109252
1
Stop Codon
range2109252
1
1174
1176
annotation2109246
1
LytC Cell Wall Binding Domain
range2109246
1
22
976
annotation2109249
1
TEV Cleavage Site
range2109249
1
1051
1104
annotation2109247
1
start
range2109247
1
22
24
annotation2109251
1
His Tag
range2109251
1
1156
1173
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
annotation1686
1
T7 TE
range1686
1
8
27
annotation1687
1
stop
range1687
1
34
34
annotation1690
1
polya
range1690
1
28
41
annotation7020
1
BBa_B0012
range7020
1
1
41
BBa_K316001
1
Pveg
pVeg Constitutive promoter for Veg locus from B. subtilis
2010-10-11T11:00:00Z
2015-05-08T01:11:56Z
PCR from existing biobrick K143053 using Pfu polymerase II
Released HQ 2013
This part is identical to the sequence submitted by Imperial 2008 team, this part was produced from K143053 by PCR.
PVeg is a constitutive promoter controlled by Sigma factor A. This promoter has two binding sites which leads to high expression of downstream genes.
There is some evidence that the sporulation master regulator the spoOA can interact with pVeg although the mechanism is not known.
false
false
_440_
0
7480
9
In stock
false
PCR using Pfu polymerase to avoid mutations
false
IC 2010 Team
annotation2085549
1
Sigma A-35
range2085549
1
63
68
annotation2085550
1
Sigma A-35
range2085550
1
86
91
BBa_K316033
1
P-RBS-CWB-
Pveg-spoVG-LytC-Flexible Linker-TEV Cleavage Site-AIP-His Tag
2010-10-25T11:00:00Z
2015-05-08T01:11:57Z
LytC cell wall binding domain <bbpart>BBa_K316030</bbpart> was extracted from ??????Bacillus subtilis?????? genome using PCR with Pfu DNA polymerase. The linker, TEV protease cleavage site and his tagged auto inducing peptide were produced by direct synthesis by mwg Eurofins[http://www.eurofinsdna.com/home.html]. The two parts were combined using AccI[http://www.neb.com/nebecomm/products/productR0161.asp] restriction endonuclease.
Introduction: This part was used to link the cell wall binding domain (CWB) of LytC <bbpart>BBa_K316030</bbpart>, with the quorum sensing peptide (AIP) as well as providing a cleavage site for a protease we want to detect. This construct was built as a protease detection unit.
LytC: The part carries part of LytC on its 5??? end. This was used to ligate the linker with LytC via an internal AccI restriction site that occurs naturally in ??????B. subtilis?????? LytC sequence.
Glycin Linker: The Linker separates the CWB and the AIP and creates space for the protease to access the cleavage site; it consists of two main sections. The first six amino acids (SRGSRA) were suggested to be used specifically with LytC<cite>1</cite> . The second section consists of a several glycin residues.
TEV Cleavage Site: This sequence forms the 3??? end of the linker and is directly attached to the 5??? end of the AIP. It is 18 amino acids (GGGGENLYFQGGKLGGGG) long and was designed to be efficiently cleaved by the TEV protease <bbpart>BBa_K316012</bbpart>, as well as being codon-optimised for expression in B. subtilis.
His-Tag: To be able to purify the protein for testing, we attached a His-Tag on our linker-AIP peptide. As it would probably interfere with recognition of the AIP by the receptor it has to be removed from the final construct.
Stop Codon: In order to end translation a double stop codon was put in place.
For more information about this part of our project please visit our wiki [http://2010.igem.org/Team:Imperial_College_London/Strategy] or take the tour[http://2010.igem.org/Team:Imperial_College_London/Tour/Page_One] to learn more about the project.
1 PMID: 14594841
(Yamamoto et al. 2003).
false
false
_440_
0
7480
9
Not in stock
false
Please refer to main text
false
IC 2010 Team
component2111917
1
BBa_B0014
component2111900
1
BBa_K316001
component2111910
1
BBa_K316043
annotation2111910
1
BBa_K316043
range2111910
1
106
1284
annotation2111917
1
BBa_B0014
range2111917
1
1293
1387
annotation2111900
1
BBa_K316001
range2111900
1
1
97
BBa_B0014_sequence
1
tcacactggctcaccttcgggtgggcctttctgcgtttatatactagagagagaatataaaaagccagattattaatccggcttttttattattt
BBa_K316001_sequence
1
aattttgtcaaaataattttattgacaacgtcttattaacgttgatataatttaaattttatttgacaaaaatgggctcgtgttgtacaataaatgt
BBa_K316043_sequence
1
tcttcagaaaggagttactagatgcgttcttatataaaagtcctaacaatgtgttttctggggctcatactttttgtgccaacagctttggccgataactcagtgaaaagagttgggggaagcaatagatacggcactgctgtacaaatatcaaagcaaatgtattcaacagcaagtacagctgtaattgttggtgggagttcctatgcagatgctatttcagcagcacctcttgcttaccagaagaatgcgccattgctttacactaattctgataagctttcatatgaaacgaaaacaagattgaaagagatgcagactaaaaatgtaattattgtaggcggaacacctgctgtttcttctaacactgctaaccagattaaaagcttggggataagtattaaacgaattgcaggaagcaaccgttatgatacggctgcacgggtggcaaaagcgatgggtgcgacttcaaaagctgttattttgaacggcttcttatatgcagacgctccggccgtcatcccttatgcagcgaaaaacgggtatccaattctttttacaaataaaacatctataaatagtgcgactacgtctgtgataaaagataagggaatttcgagtaccgttgttgtaggaggcactggaagtatcagcaatacggtatacaacaagttaccttctcctacaagaattagcggttcaaacagatatgagcttgctgcaaatatcgtacaaaaacttaatttatcaacaagcaccgtatatgtaagcaatggattcagctaccctgactctattgcaggagctacactggcagctaagaagaagcaatctcttattcttacaaatggtgaaaatttatctacaggagcccgtaaaattattggaagtaaaaacatgtcaaactttatgattatcggaaacactcctgccgtaagcacaaaggttgctaatcagctaaagaatccagttgtaagtagaggatcccgcgctctgggaggcggcggctcaggtggtggcggaagcggcggaggcggttctgccgcagcgggaggtggaggtgaaaatctttattttcaaggaggcaaactgggaggcggcggagaaatgcgtctgtcaaaattctttcgcgattttattcttcaaagaaaaaagcatcaccatcatcaccattaataa
BBa_K316033_sequence
1
aattttgtcaaaataattttattgacaacgtcttattaacgttgatataatttaaattttatttgacaaaaatgggctcgtgttgtacaataaatgttactagagtcttcagaaaggagttactagatgcgttcttatataaaagtcctaacaatgtgttttctggggctcatactttttgtgccaacagctttggccgataactcagtgaaaagagttgggggaagcaatagatacggcactgctgtacaaatatcaaagcaaatgtattcaacagcaagtacagctgtaattgttggtgggagttcctatgcagatgctatttcagcagcacctcttgcttaccagaagaatgcgccattgctttacactaattctgataagctttcatatgaaacgaaaacaagattgaaagagatgcagactaaaaatgtaattattgtaggcggaacacctgctgtttcttctaacactgctaaccagattaaaagcttggggataagtattaaacgaattgcaggaagcaaccgttatgatacggctgcacgggtggcaaaagcgatgggtgcgacttcaaaagctgttattttgaacggcttcttatatgcagacgctccggccgtcatcccttatgcagcgaaaaacgggtatccaattctttttacaaataaaacatctataaatagtgcgactacgtctgtgataaaagataagggaatttcgagtaccgttgttgtaggaggcactggaagtatcagcaatacggtatacaacaagttaccttctcctacaagaattagcggttcaaacagatatgagcttgctgcaaatatcgtacaaaaacttaatttatcaacaagcaccgtatatgtaagcaatggattcagctaccctgactctattgcaggagctacactggcagctaagaagaagcaatctcttattcttacaaatggtgaaaatttatctacaggagcccgtaaaattattggaagtaaaaacatgtcaaactttatgattatcggaaacactcctgccgtaagcacaaaggttgctaatcagctaaagaatccagttgtaagtagaggatcccgcgctctgggaggcggcggctcaggtggtggcggaagcggcggaggcggttctgccgcagcgggaggtggaggtgaaaatctttattttcaaggaggcaaactgggaggcggcggagaaatgcgtctgtcaaaattctttcgcgattttattcttcaaagaaaaaagcatcaccatcatcaccattaataatactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagagagaatataaaaagccagattattaatccggcttttttattattt
BBa_B0011_sequence
1
agagaatataaaaagccagattattaatccggcttttttattattt
BBa_B0012_sequence
1
tcacactggctcaccttcgggtgggcctttctgcgtttata
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