BBa_K1088000
1
dxs (B. su
1-deoxyxylulose-5-phosphate synthase (Dxs) enzyme from MEP pathway isolated from B. subtilis 168
2013-07-08T11:00:00Z
2015-05-08T01:09:06Z
B. subtilis 168
The MEP pathway is a common bacterial isoprenoid pathway. To improve the yield of the end product, isopentenyl diphosphate (IPP) and dimethylallyl pyrophosphate(DMAPP), Zhao et al. expressed, in 2011, the 1-deoxyxylulose-5-phosphate synthase(Dxs)gene aswell as the second step of the MEP pathway. They saw a 2.3 fold increase in isoprene output (isoprene is one of many products from the MEP pathway).
From Biocyc:
"Dxs catalyzes the thiamin diphosphate-dependent reaction that condenses pyruvate and D-glyceraldehyde-3-phosphate to yield 1-deoxy-D-xylulose 5-phosphate [Julsing07]. This is the first, rate-limiting step in the methylerythritol phosphate pathway of isoprenoid biosynthesis, and is believed to feed into the pyridoxal 5-phosphate (vitamin B6) and thiamin (vitamin B1) biosynthesis pathways."
-http://biocyc.org/BSUB/NEW-IMAGE?type=ENZYME&object=BSU24270-MONOMER
false
false
_1398_
0
12807
9
It's complicated
false
we changed TTG start codon to ATG to improve translation efficiency.
We performed 4 silent site directed mutageneses to remove 2 EcoR1 and 2 Pst1 restriction sites
false
Andreas Kj??r
annotation2331286
1
Dxs B. sub coding region
range2331286
1
1
1901
BBa_K592009
1
amilCP
amilCP, blue chromoprotein
2011-09-17T11:00:00Z
2015-05-08T01:12:48Z
Acropora millepora
Released HQ 2013
This chromoprotein, amilCP, naturally exhibits very strong color when expressed. The color is blue/purple and is visible to naked eye, thereby requiring no instruments to observe. This DNA was provided by Jeffrey Miller at UCLA. It was made BioBrick-compatible after removal of one illegal internal restriction site (EcoRI).
false
false
_763_
0
7929
9
In stock
true
Illegal internal restriction site had to be removed (EcoRI).
false
Lei Sun
annotation2131628
1
amilCP
range2131628
1
1
666
BBa_J15001
1
BBa_J15001
strong synthetic E. coli ribosome binding site with SacI site.
2007-07-12T11:00:00Z
2015-08-31T04:08:32Z
Synthetic.
This is a strong synthetic E. coli ribosome binding site. It is synthesised as two complementary oligonucleotides rather than being incorporated into a biobrick plasmid. It incorporates a SacI site overlapping the XbaI site, which is preserved when it is added to any other biobrick. This allows easy detection of the RBS after it has been added upstream of a biobrick coding sequence in a plasmid vector.
false
false
_163_
0
837
163
Not in stock
false
Note the presence of a SacI site overlapping the XbaI site, which is preserved when this biobrick is added to any other biobrick. At the time of writing, this biobrick is added as a short piece of DNA composed of two complementary oligonucleotides rather than being incorporated into a biobrick cloning vector by itself. It can be added upstream of a biobrick coding sequence, and its presence can easily be detected in miniprep DNA on a gel by using a SacI-SpeI or similar digest.
false
Chris French
annotation1938046
1
rbs
range1938046
1
4
10
annotation1938045
1
SacI
range1938045
1
1
3
BBa_K1088006
1
BBa_K1088006
B. subtilis dxs-AmilCP fusion (constitutive promoter)
2013-08-19T11:00:00Z
2015-05-08T01:09:06Z
The parts used in this composite parts derives mainly from E. coli, however the amilCP derives from Acropora millepora.
This composite part consists of 5 sub parts
A strong constitutive promoter: BBa_J23100
A strong RBS: BBa_J15001
The coding region of Dxs from B. sub: BBa_K1088000
A 10 aa Protein Linker: BBa_K105012
And the AmilCP chromoprotein: BBa_K592009
The idea was to detect expression of dxs protein by the reporter amilCP fused via a linker to the Dxs protein. Unfortunately it seems as though the amilCP protein does not work well as a protein-fusion reporter, and the color change was vague.
false
false
_1398_
0
12807
9
It's complicated
false
We added the linker, because we hoped it would improve the function of the chromoprotein amilCP. However it seems as though this did not work as intended. The promoter and RBS are as strong as we could find them, since we only where interested in knowing if the Dxs where expressed at all.
false
Andreas Kj??r
component2373520
1
BBa_J23100
component2373528
1
BBa_K592009
component2373523
1
BBa_J15001
component2373526
1
BBa_K105012
component2373525
1
BBa_K1088000
annotation2373526
1
BBa_K105012
range2373526
1
1948
1977
annotation2373525
1
BBa_K1088000
range2373525
1
46
1947
annotation2373520
1
BBa_J23100
range2373520
1
1
35
annotation2373528
1
BBa_K592009
range2373528
1
1978
2646
annotation2373523
1
BBa_J15001
range2373523
1
36
45
BBa_J23100
1
BBa_J23100
constitutive promoter family member
2006-08-03T11:00:00Z
2015-08-31T04:08:40Z
Isolated from library of promoters
Released HQ 2013
Replace later
false
true
_52_
0
483
95
In stock
true
N/A
true
John Anderson
BBa_K105012
1
BBa_K105012
10 aa flexible protein domain linker
2008-10-18T11:00:00Z
2015-05-08T01:08:52Z
Oligos with a yeast optimized coding for the peptide GENLYFQSGG have been ordered
<br\n>
The amino acide sequence corresponds to the interdomaine linker of xxx.
''paper''
This is a 10 amino acides long linker peptide which can be used to join protein domains together in a flexible way. So fusion proteins with variable DNA-binding and activation or repression domains might be assembled.
<br\n>
false
true
_253_
0
3313
9
In stock
true
To enable the fusion with other domains in frame the vector of this BioBrick has no base pair in between the restriction side and the BioBrick. Furthermore, this coding sequence does not include a start codon.<br\n>
For more information about this issus, see:<br\n>
Phillips, I.E. and Silver, P.A. "A new Biobrick Assembly Strategy Designed for Facile Protein Engineering." <br\n>
DSpace http://hdl.handle.net/1721.1/32535 (2006).
true
Manuel Gersbacher, Katja Karstens
BBa_K1088006_sequence
1
ttgacggctagctcagtcctaggtacagtgctagcctcaaggaggatggatcttttatcaatacaggacccgtcgtttttaaaaaacatgtccattgatgaattagagaaattaagtgatgaaatccgtcagtttttaattacaagtttatccgcttccggcggccacatcggcccaaacttaggtgtcgtagagcttactgttgccctgcataaggaatttaacagcccgaaagacaaatttttatgggatgtaggccatcagtcgtatgtccataagctgctgacaggacgcggaaaagaatttgcgacgcttcgccagtacaaagggctttgcggatttccaaagcggagtgaaagcgagcacgatgtttgggaaaccgggcacagctcgacttctctgtcaggcgcgatgggaatggcagctgcccgtgatattaaaggaacggatgaatatattattccgatcattggtgacggcgcgctgaccggcggtatggcgctcgaagcccttaaccacatcggcgacgagaaaaaagacatgattgtcatccttaatgataatgaaatgagtattgcgccaaacgtcggtgccattcactctatgctcggacggctccgcactgcggggaaataccagtgggtcaaagatgagcttgaatacttatttaaaaagattccggcagttgggggcaagcttgccgccacggcggaacgggtcaaagacagcctgaaatacatgctcgtctccggaatgtttttcgaggagctcggttttacgtatttgggcccagtggacggacattcttatcatgagctgattgagaatcttcaatacgccaaaaaaacgaaaggccctgttcttctgcacgtcattacgaaaaaagggaaggggtacaaaccggctgagaccgatacgattgggacatggcatggtaccggaccatataaaattaataccggtgactttgtaaagccgaaagccgcagctccttcgtggagcggtcttgtcagcggaactgtgcagcgaatggcgcgcgaggacggacgcattgtagccattacgccggctatgcctgtcggttcaaagcttgaaggcttcgcaaaggaattccctgaccggatgttcgacgtaggaatcgcagaacagcatgccgcaacaatggctgcagctatggcaatgcagggtatgaagccgtttttggcgatttactcaaccttcctgcaaagggcatatgaccaagttgttcatgacatctgccgccaaaacgctaatgtgtttattggaattgaccgtgctggactcgttggcgctgatggagagacacatcaaggcgtgtttgatattgcgtttatgcgccacattccaaacatggtcttaatgatgccgaaagacgaaaatgaaggccagcacatggttcatacagcacttagctatgacgaaggcccgatagcaatgcgttttccgcgcggaaacggactcggcgtaaaaatggatgaacagttgaaaacgattccgatcggtacgtgggaggtgctgcgtccagggaacgatgctgtcatcttaacattcggcacaacaatcgaaatggcgattgaagcagccgaagagctgcagaaagaaggcctttccgtgcgcgttgtgaatgcgcgttttattaagccgattgatgaaaagatgatgaagagtatcctaaaagaaggcttgccaattttaacaattgaagaagcggtcttagaaggcggtttcggaagctcgattttagaattcgctcatgatcaaggtgaatatcatactccgattgacagaatgggtatacctgatcggtttattgaacacggaagtgtaacagcgcttcttgaggaaattggactgacaaaacagcaggtggcaaatcgtattagattactgatgccaccaaagacacacaaaggaattggatcatgaggtgaaaatttgtattttcaatctggtggtatgagtgtgatcgctaaacaaatgacctacaaggtttatatgtcaggcacggtcaatggacactactttgaggtcgaaggcgatggaaaaggtaagccctacgagggggagcagacggtaaagctcactgtcaccaagggcggacctctgccatttgcttgggatattttatcaccacagtgtcagtacggaagcataccattcaccaagtaccctgaagacatccctgactatgtaaagcagtcattcccggagggctatacatgggagaggatcatgaactttgaagatggtgcagtgtgtactgtcagcaatgattccagcatccaaggcaactgtttcatctaccatgtcaagttctctggtttgaactttcctcccaatggacctgtcatgcagaagaagacacagggctgggaacccaacactgagcgtctctttgcacgagatggaatgctgctaggaaacaactttatggctctgaagttagaaggaggcggtcactatttgtgtgaatttaaaactacttacaaggcaaagaagcctgtgaagatgccagggtatcactatgttgaccgcaaactggatgtaaccaatcacaacaaggattacacttcggttgagcagtgtgaaatttccattgcacgcaaacctgtggtcgcctaataa
BBa_J23100_sequence
1
ttgacggctagctcagtcctaggtacagtgctagc
BBa_J15001_sequence
1
ctcaaggagg
BBa_K592009_sequence
1
atgagtgtgatcgctaaacaaatgacctacaaggtttatatgtcaggcacggtcaatggacactactttgaggtcgaaggcgatggaaaaggtaagccctacgagggggagcagacggtaaagctcactgtcaccaagggcggacctctgccatttgcttgggatattttatcaccacagtgtcagtacggaagcataccattcaccaagtaccctgaagacatccctgactatgtaaagcagtcattcccggagggctatacatgggagaggatcatgaactttgaagatggtgcagtgtgtactgtcagcaatgattccagcatccaaggcaactgtttcatctaccatgtcaagttctctggtttgaactttcctcccaatggacctgtcatgcagaagaagacacagggctgggaacccaacactgagcgtctctttgcacgagatggaatgctgctaggaaacaactttatggctctgaagttagaaggaggcggtcactatttgtgtgaatttaaaactacttacaaggcaaagaagcctgtgaagatgccagggtatcactatgttgaccgcaaactggatgtaaccaatcacaacaaggattacacttcggttgagcagtgtgaaatttccattgcacgcaaacctgtggtcgcctaataa
BBa_K1088000_sequence
1
atggatcttttatcaatacaggacccgtcgtttttaaaaaacatgtccattgatgaattagagaaattaagtgatgaaatccgtcagtttttaattacaagtttatccgcttccggcggccacatcggcccaaacttaggtgtcgtagagcttactgttgccctgcataaggaatttaacagcccgaaagacaaatttttatgggatgtaggccatcagtcgtatgtccataagctgctgacaggacgcggaaaagaatttgcgacgcttcgccagtacaaagggctttgcggatttccaaagcggagtgaaagcgagcacgatgtttgggaaaccgggcacagctcgacttctctgtcaggcgcgatgggaatggcagctgcccgtgatattaaaggaacggatgaatatattattccgatcattggtgacggcgcgctgaccggcggtatggcgctcgaagcccttaaccacatcggcgacgagaaaaaagacatgattgtcatccttaatgataatgaaatgagtattgcgccaaacgtcggtgccattcactctatgctcggacggctccgcactgcggggaaataccagtgggtcaaagatgagcttgaatacttatttaaaaagattccggcagttgggggcaagcttgccgccacggcggaacgggtcaaagacagcctgaaatacatgctcgtctccggaatgtttttcgaggagctcggttttacgtatttgggcccagtggacggacattcttatcatgagctgattgagaatcttcaatacgccaaaaaaacgaaaggccctgttcttctgcacgtcattacgaaaaaagggaaggggtacaaaccggctgagaccgatacgattgggacatggcatggtaccggaccatataaaattaataccggtgactttgtaaagccgaaagccgcagctccttcgtggagcggtcttgtcagcggaactgtgcagcgaatggcgcgcgaggacggacgcattgtagccattacgccggctatgcctgtcggttcaaagcttgaaggcttcgcaaaggaattccctgaccggatgttcgacgtaggaatcgcagaacagcatgccgcaacaatggctgcagctatggcaatgcagggtatgaagccgtttttggcgatttactcaaccttcctgcaaagggcatatgaccaagttgttcatgacatctgccgccaaaacgctaatgtgtttattggaattgaccgtgctggactcgttggcgctgatggagagacacatcaaggcgtgtttgatattgcgtttatgcgccacattccaaacatggtcttaatgatgccgaaagacgaaaatgaaggccagcacatggttcatacagcacttagctatgacgaaggcccgatagcaatgcgttttccgcgcggaaacggactcggcgtaaaaatggatgaacagttgaaaacgattccgatcggtacgtgggaggtgctgcgtccagggaacgatgctgtcatcttaacattcggcacaacaatcgaaatggcgattgaagcagccgaagagctgcagaaagaaggcctttccgtgcgcgttgtgaatgcgcgttttattaagccgattgatgaaaagatgatgaagagtatcctaaaagaaggcttgccaattttaacaattgaagaagcggtcttagaaggcggtttcggaagctcgattttagaattcgctcatgatcaaggtgaatatcatactccgattgacagaatgggtatacctgatcggtttattgaacacggaagtgtaacagcgcttcttgaggaaattggactgacaaaacagcaggtggcaaatcgtattagattactgatgccaccaaagacacacaaaggaattggatcatga
BBa_K105012_sequence
1
ggtgaaaatttgtattttcaatctggtggt
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