BBa_M36282
1
BBa_M36282
5' Bicistronic UTR, includes ATG start codon
2011-11-29T12:00:00Z
2015-05-08T01:14:04Z
BioFab: apFAB563
This sequence contains an RBS followed by the sequence TAATG. The parts number is BBa_M36000 (PartsRegistry). The first RBS sequence is strong, while the second one is weak. This stops the first polymerase with the stop codon TAA, but allows for a second polymerase to bind to the start codon ATG(BIOFAB). This sequence prevents a hairpin loop from being formed with another part of the sequence, which would decrease the amount of translation. The bicistronic sequence ends with a start codon ATG.
false
true
_848_
0
11068
9
Not in stock
false
no
false
Sunil Bodapati, Julia Joung, Daniel Esquivel-Reynoso
annotation2166677
1
Start Codon
range2166677
1
86
88
BBa_M36281
1
BBa_M36281
High Efficiency Terminator
2011-11-29T12:00:00Z
2015-05-08T01:14:04Z
BIOFAB: apFAB391
This terminator works with 99% efficiency.
false
false
_848_
0
11068
9
Not in stock
false
High efficiency terminator
false
Sunil Bodapati, Julia Joung, Daniel Esquivel-Reynoso
BBa_M36283
1
BBa_M36283
Human Alcohol Dehydrogenase 1B actuator
2011-11-29T12:00:00Z
2015-05-08T01:14:04Z
Bicistonic RBS - BioFab: apFAB563
ADH1B - partsregistry: BBa_M36666
Histidine Tag - 6 histidines
Stop Codon - generic TAG
Terminator - BIOFAB: apFAB391
Our proposed gene sequence is an actuator that produces ADH1B following an RNA polymerase per second (PoPS) signal from a sensor. The sensor will be followed by a bi-cistronic ribosome binding sequence provided by BIOFAB (BD2). This type of ribosome binding sequence contains a ribosome binding site followed by the DNA sequence TAATG, which terminates translation of the first part with stop codon TAA. A second ribosome can bind in the sequence and start translation of the gene with start codon ATG. The bi-cistronic ribosome binding sequence prevents the protein???s ribosome binding site from forming a hairpin loop with another part of the sequence, which would reduce the amount of translation. Specifically, our group chose to use BD2 because it offers consistently high to medium expression of the gene of interest. After BD2, we have inserted the gene sequence of ADH1B, which we constructed using the protein sequence from protein database UniProt that was cross referenced with NCBI. Since we are inserting a eukaryotic gene into a prokaryote, we optimized the DNA sequence for E. Coli using a codon optimizing chart provided by Sharp et al. (1988), selecting for highly transcribed genes as our objective is to produce large amounts of the enzyme. Our ADH1B gene was followed by a histidine tag consisting of 6 histidines that can be used to purify the enzyme with nickel affinity chromatography. Then we put a stop codon TAG that terminates translation. Lastly, we placed a transcription terminator, part apFAB391 provided by Biofab, that terminates transcription 99% of the time because our objective is to transcribe ADH1B so we would want to prevent transcribing other proteins on the same plasmid to conserve energy.
true
false
_848_
0
11068
9
Discontinued
false
We maximized expression by using a highly expressing bicistronic RBS. We needed a way to purify the enzyme post production. The histidine tag allows us to perform nickel affinity chromatography to isolate purified protein.
false
Sunil Bodapati, Julia Joung, Daniel Esquivel-Reynoso
component2166159
1
BBa_M36281
component2166158
1
BBa_M36280
component2166160
1
BBa_M36282
annotation2166158
1
BBa_M36280
range2166158
1
1
1143
annotation2166159
1
BBa_M36281
range2166159
1
1152
1233
annotation2166160
1
BBa_M36282
range2166160
1
1242
1329
BBa_M36280
1
BBa_M36280
Human Alcohol Dehydrogenase 1B
2011-11-29T12:00:00Z
2015-05-08T01:14:04Z
P00325 (ADH1B_HUMAN) provided by UniProt.
This gene sequence encodes for the human protein alcohol dehydrogenase which converts ethanol into acetaldehyde in the presence of zinc as its cofactor.
false
false
_848_
0
11068
9
Not in stock
false
This sequence has been codon optimized for high expression in E. Coli.
false
Sunil Bodapati, Julia Joung, Daniel Esquivel-Reynoso
annotation2166154
1
Histidine
range2166154
1
1123
1140
annotation2166155
1
stop codon
range2166155
1
1141
1143
annotation2166488
1
alcohol dehydrogenase 1b
range2166488
1
1
1122
BBa_M36280_sequence
1
tctactgctggtaaagttatcaaatgcaaagctgctgttctgtgggaagttaaaaaaccgttctctatcgaagacgttgaagttgctccgccgaaagcttacgaagttcgtatcaaaatggttgctgttggtatctgccgtactgacgaccacgttgtttctggtaacctggttactccgctgccggttatcctgggtcacgaagctgctggtatcgttgaatctgttggtgaaggtgttactactgttaaaccgggtgacaaagttatcccgctgttcactccgcagtgcggtaaatgccgtgtttgcaaaaacccggaatctaactactgcctgaaaaacgacctgggtaacccgcgtggtactctgcaggacggtactcgtcgtttcacttgccgtggtaaaccgatccaccacttcctgggtacttctactttctctcagtacactgttgttgacgaaaacgctgttgctaaaatcgacgctgcttctccgctggaaaaagtgtgcctgatcggttgcggtttctctactggttacggttccgctgttaacgttgctaaagttactccgggttctacttgcgctgttttcggtctgggtggtgttggtctgtctgctgttatgggttgcaaagcagctggcgctgctcgtatcatcgctgtggacatcaacaaagacaaattcgctaaagctaaagaactgggtgctactgaatgcatcaacccgcaggactacaaaaaaccgatccaggaagttctgaaagaaatgactgacggtggcgttgacttctcgttcgaagttatcggtcgtctggacactatgatggcttctctgctgtgctgccacgaagcatgcggcacctccgtaatcgttggtgttccgccggcttcccagaacctgtccatcaacccgatgctgctgctgaccggccgtacttggaaaggtgctgtttacggcggcttcaaatctaaagaaggtatcccgaaactggtagctgacttcatggcaaaaaaattctccctggacgctctgatcacccacgttctgccgttcgaaaaaatcaacgaaggtttcgacctgctgcactccggcaaatctatccgtactgttctgaccttccaccaccatcaccaccattag
BBa_M36282_sequence
1
gggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgctaaggaggttttctaatg
BBa_M36281_sequence
1
tcggtcagtttcacctgatttacgtaaaaacccgcttcggcgggtttttgcttttggaggggcagaaagatgaatgactgtc
BBa_M36283_sequence
1
tctactgctggtaaagttatcaaatgcaaagctgctgttctgtgggaagttaaaaaaccgttctctatcgaagacgttgaagttgctccgccgaaagcttacgaagttcgtatcaaaatggttgctgttggtatctgccgtactgacgaccacgttgtttctggtaacctggttactccgctgccggttatcctgggtcacgaagctgctggtatcgttgaatctgttggtgaaggtgttactactgttaaaccgggtgacaaagttatcccgctgttcactccgcagtgcggtaaatgccgtgtttgcaaaaacccggaatctaactactgcctgaaaaacgacctgggtaacccgcgtggtactctgcaggacggtactcgtcgtttcacttgccgtggtaaaccgatccaccacttcctgggtacttctactttctctcagtacactgttgttgacgaaaacgctgttgctaaaatcgacgctgcttctccgctggaaaaagtgtgcctgatcggttgcggtttctctactggttacggttccgctgttaacgttgctaaagttactccgggttctacttgcgctgttttcggtctgggtggtgttggtctgtctgctgttatgggttgcaaagcagctggcgctgctcgtatcatcgctgtggacatcaacaaagacaaattcgctaaagctaaagaactgggtgctactgaatgcatcaacccgcaggactacaaaaaaccgatccaggaagttctgaaagaaatgactgacggtggcgttgacttctcgttcgaagttatcggtcgtctggacactatgatggcttctctgctgtgctgccacgaagcatgcggcacctccgtaatcgttggtgttccgccggcttcccagaacctgtccatcaacccgatgctgctgctgaccggccgtacttggaaaggtgctgtttacggcggcttcaaatctaaagaaggtatcccgaaactggtagctgacttcatggcaaaaaaattctccctggacgctctgatcacccacgttctgccgttcgaaaaaatcaacgaaggtttcgacctgctgcactccggcaaatctatccgtactgttctgaccttccaccaccatcaccaccattagtactagagtcggtcagtttcacctgatttacgtaaaaacccgcttcggcgggtttttgcttttggaggggcagaaagatgaatgactgtctactagaggggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgctaaggaggttttctaatg
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