BBa_I742144
1
Plac-sam8
Plac+lacZ+sam8 (tyrosine ammonia lyase)
2007-10-22T11:00:00Z
2015-08-31T04:08:03Z
sam8 is derived from Saccharothix espanaensis DSM 44229. The lac promoter and lacZ' minigene, encoding the N-terminal 76 amino acid residues of LacZ, are derived from Escherichia coli BS(DE3).
This part consists of sam8, encoding tyrosine ammonia lyase (converts tyrosine to p-coumaric acid) from the bacterium Saccharothrix espanaensis DSM 44229, inserted downstream of a lac promoter with attached lacZ' minigene. The purpose of this was to ensure sam8 expression on induction with IPTG and also to provide a visually selectable tag when assembling the entire pathway; by always inserting a lacZ'-containing part into a non-lacZ'-containing part it is easy to select the correct clones on the basis that they form blue colonies.
false
false
_123_
0
837
163
It's complicated
true
No important considerations.
false
Chris French
component2219402
1
BBa_I742148
component2219399
1
BBa_J33207
annotation2219399
1
BBa_J33207
range2219399
1
1
600
annotation2219402
1
BBa_I742148
range2219402
1
609
2158
BBa_I742146
1
BBa_I742146
sam8 native rbs
2007-10-23T11:00:00Z
2015-08-31T04:08:03Z
Saccharothrix espanaensis DSM 44229.
Native ribosome binding site of sam8 (tyrosine ammonia lyase) from the bacterium Saccharothrix espanaensis DSM 44229.
false
false
_123_
0
837
163
Not in stock
false
Virtual part to be added to coding sequence I742142.
false
Chris French
BBa_J33207
1
BBa_J33207
lac promoter and lacZ
2006-10-26T11:00:00Z
2015-08-31T04:08:46Z
The DNA was amplified from E. coli BL21 genomic DNA using primers based on published sequence (Genbank accession J01636, gi:146575). The annotation shown here is based on that associated with this Genbank entry. The sequence shown here is derived by sequencing the construct.
This part (submitted in pSB1A2) consists of the lac promoter and lacZ' gene, encoding the N-terminal 76 amino acid residues of LacZ, sufficient to complement the lacZ-delta-M15 mutation for blue-white selection on Xgal plates. A SacI site has been introduced at the 3' end, overlapping the XbaI site of the Biobrick prefix. This is designed to be used as a cloning vector for making new biobricks. PCR primers can be designed with a SacI site in one primer and an SpeI site in the other. This removes the necessity for an excessively long non-complementary tail on one primer bearing either the full biobrick prefix or suffix. The PCR product can then be digested with SacI and SpeI for insertion into this plasmid, replacing the Plac-lacZ' cassette. Recombinant plasmids will then be white on IPTG/Xgal plates, whereas any that still contain the original insert will be blue. We have used this strategy to prepare several biobricks, including BBa_J33204, which contains the xylE gene encoding catechol-2,3-dioxygenase. (For making biobricks that contain lacZ', BBa_J33204 can be used in the same way; in this case, clones with plasmids that still contain xylE will turn yellow on addition of a drop of 10 mM catechol.)
false
false
_63_
0
837
63
It's complicated
true
Note that the SacI site overlaps the SpeI site. The Biobrick prefix ends ...TCTAGAG. When this is added to the CTC at the start of the sequence shown here, the SacI site, GAGCTC, is generated.
false
Chris French
annotation1907855
1
CAP binding site
range1907855
1
248
285
annotation1907859
1
-10
range1907859
1
320
325
annotation1907858
1
lacZ'
range1907858
1
370
600
annotation1907854
1
SacI
range1907854
1
1
3
annotation1907857
1
rbs
range1907857
1
359
362
annotation1907860
1
-35
range1907860
1
297
302
annotation1907856
1
LacI binding site
range1907856
1
332
352
BBa_I742142
1
sam8
sam8 (tyrosine-ammonia lyase) coding sequence
2007-10-22T11:00:00Z
2015-08-31T04:08:03Z
Saccharothrix espanaensis DSM 4429. Accession: ABC88666.
This is the coding sequence of sam8, encoding tyrosine ammonia lyase, from the bacterium Saccharothrix espanaensis DSM 4429. The enzyme converts tyrosine to p-coumaric acid. Accession: ABC88666. Reference: Berner, M., Krug, D., Bihlmaier, C., Vente, A., Muller, R. and Bechthold, A. 'Genes and Enzymes Involved in Caffeic Acid Biosynthesis in the Actinomycete Saccharothrix espanaensis' J. Bacteriol. 188 (7), 2666-2673 (2006)
false
false
_123_
0
837
163
It's complicated
true
No important considerations.
false
sarah hollingshead
BBa_I742148
1
rbs+sam8
sam8 (tyrosine ammonia lyase) with native rbs
2007-10-23T11:00:00Z
2015-08-31T04:08:03Z
Saccharothrix espanaensis DSM 44229
This is sam8 (tyrosine ammonia lyase) of the bacterium Saccharothrix espanaensis DSM 44229 with its native ribosome binding site.
false
false
_123_
0
837
163
It's complicated
true
This part was made as a single biobrick rather than separate rbs and coding sequence, hence the use of blunt ligation in generating the final sequence.
false
sarah hollingshead
component1953823
1
BBa_I742146
component1953824
1
BBa_I742142
annotation1953823
1
BBa_I742146
range1953823
1
1
14
annotation1953824
1
BBa_I742142
range1953824
1
15
1550
BBa_I742142_sequence
1
atgacgcaggtcgtggaacgtcaggctgatcggctcagcagcagggagtacctggcccgggtcgtgcgcagcgccgggtgggacgccggtctcacctcgtgcaccgacgaggagatcgtccggatgggcgcgagcgcgcgcaccatcgaggagtacctgaagtccgacaagcccatctacggcctgacgcagggcttcggtccgctggtgctgttcgacgccgactcggagctggagcagggcggctcgctgatctcgcacctgggcaccggccagggcgcgccactggccccggaggtgtcgcggctgatcctctggctgcgcatccagaacatgcgcaaggggtactcggcggtctcgccggtgttctggcagaagctcgccgacctgtggaacaaggggttcaccccggcgatcccccggcacggcacggtcagcgcgagcggcgacctgcaaccgctggcgcacgccgcgctcgccttcaccggtgtcggcgaggcgtggacccgggacgccgacggccggtggtccaccgtgccggccgtggacgcgctcgccgcgctgggggcggagccgttcgactggccggtgcgcgaggcgctggcgttcgtcaacgggaccggcgcgagcctcgcggtggctgtgctcaaccaccggtccgccctgcggctggtccgcgcctgcgccgtgctctccgcgcggctggcgaccctgctgggggccaatcccgagcactacgacgtggggcacggtgtcgcgcgcggccaggtcggtcagctgaccgcggcggagtggatccggcaggggctgccccggggcatggtgcgcgacggcagccgcccgctccaggagccgtacagcctgcggtgcgcgccgcaggtgctcggcgcggtgctcgaccagctcgacggcgcgggcgacgtgctggcgcgggaggtcgacggctgccaggacaacccgatcacctacgagggcgagctgctgcacggcggcaacttccacgccatgccggtgggtttcgcctccgaccagatcgggttggccatgcacatggccgcctacctggccgagcgccagctgggtctgctggtcagcccggtgaccaacggcgacctgccgcccatgctcaccccgcgcgccgggcgcggtgccgggctggccggggtgcagatcagcgcgacctcgttcgtctcgcggatccggcagctggtgttccccgcctcgctgaccaccctgccgaccaacggctggaaccaggaccacgtgccgatggcgctcaacggggcgaactcggtgttcgaggcgttggagctcggctggctgacggtcgggtcgctggcggtgggcgtcgcgcagctcgcggccatgaccggccacgccgcggagggcgtctgggcggagctggccgggatctgcccgccgctggacgccgaccgcccgctgggcgccgaggtgcgcgccgcgcgcgacctgctgtccgcgcacgcggaccaactgctcgtcgacgaggcagacgggaaggatttcggataataa
BBa_I742144_sequence
1
ctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtgaaattgtgagcggataacaatttcacacaggaaacagctatgaccatgattacggattcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggcgctttgcctggtttccggcaccagaagcggtgccggaaagctggctggagtgatactagaggaggttgggacaagatgacgcaggtcgtggaacgtcaggctgatcggctcagcagcagggagtacctggcccgggtcgtgcgcagcgccgggtgggacgccggtctcacctcgtgcaccgacgaggagatcgtccggatgggcgcgagcgcgcgcaccatcgaggagtacctgaagtccgacaagcccatctacggcctgacgcagggcttcggtccgctggtgctgttcgacgccgactcggagctggagcagggcggctcgctgatctcgcacctgggcaccggccagggcgcgccactggccccggaggtgtcgcggctgatcctctggctgcgcatccagaacatgcgcaaggggtactcggcggtctcgccggtgttctggcagaagctcgccgacctgtggaacaaggggttcaccccggcgatcccccggcacggcacggtcagcgcgagcggcgacctgcaaccgctggcgcacgccgcgctcgccttcaccggtgtcggcgaggcgtggacccgggacgccgacggccggtggtccaccgtgccggccgtggacgcgctcgccgcgctgggggcggagccgttcgactggccggtgcgcgaggcgctggcgttcgtcaacgggaccggcgcgagcctcgcggtggctgtgctcaaccaccggtccgccctgcggctggtccgcgcctgcgccgtgctctccgcgcggctggcgaccctgctgggggccaatcccgagcactacgacgtggggcacggtgtcgcgcgcggccaggtcggtcagctgaccgcggcggagtggatccggcaggggctgccccggggcatggtgcgcgacggcagccgcccgctccaggagccgtacagcctgcggtgcgcgccgcaggtgctcggcgcggtgctcgaccagctcgacggcgcgggcgacgtgctggcgcgggaggtcgacggctgccaggacaacccgatcacctacgagggcgagctgctgcacggcggcaacttccacgccatgccggtgggtttcgcctccgaccagatcgggttggccatgcacatggccgcctacctggccgagcgccagctgggtctgctggtcagcccggtgaccaacggcgacctgccgcccatgctcaccccgcgcgccgggcgcggtgccgggctggccggggtgcagatcagcgcgacctcgttcgtctcgcggatccggcagctggtgttccccgcctcgctgaccaccctgccgaccaacggctggaaccaggaccacgtgccgatggcgctcaacggggcgaactcggtgttcgaggcgttggagctcggctggctgacggtcgggtcgctggcggtgggcgtcgcgcagctcgcggccatgaccggccacgccgcggagggcgtctgggcggagctggccgggatctgcccgccgctggacgccgaccgcccgctgggcgccgaggtgcgcgccgcgcgcgacctgctgtccgcgcacgcggaccaactgctcgtcgacgaggcagacgggaaggatttcggataataa
BBa_I742146_sequence
1
gaggttgggacaag
BBa_J33207_sequence
1
ctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtgaaattgtgagcggataacaatttcacacaggaaacagctatgaccatgattacggattcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggcgctttgcctggtttccggcaccagaagcggtgccggaaagctggctggagtga
BBa_I742148_sequence
1
gaggttgggacaagatgacgcaggtcgtggaacgtcaggctgatcggctcagcagcagggagtacctggcccgggtcgtgcgcagcgccgggtgggacgccggtctcacctcgtgcaccgacgaggagatcgtccggatgggcgcgagcgcgcgcaccatcgaggagtacctgaagtccgacaagcccatctacggcctgacgcagggcttcggtccgctggtgctgttcgacgccgactcggagctggagcagggcggctcgctgatctcgcacctgggcaccggccagggcgcgccactggccccggaggtgtcgcggctgatcctctggctgcgcatccagaacatgcgcaaggggtactcggcggtctcgccggtgttctggcagaagctcgccgacctgtggaacaaggggttcaccccggcgatcccccggcacggcacggtcagcgcgagcggcgacctgcaaccgctggcgcacgccgcgctcgccttcaccggtgtcggcgaggcgtggacccgggacgccgacggccggtggtccaccgtgccggccgtggacgcgctcgccgcgctgggggcggagccgttcgactggccggtgcgcgaggcgctggcgttcgtcaacgggaccggcgcgagcctcgcggtggctgtgctcaaccaccggtccgccctgcggctggtccgcgcctgcgccgtgctctccgcgcggctggcgaccctgctgggggccaatcccgagcactacgacgtggggcacggtgtcgcgcgcggccaggtcggtcagctgaccgcggcggagtggatccggcaggggctgccccggggcatggtgcgcgacggcagccgcccgctccaggagccgtacagcctgcggtgcgcgccgcaggtgctcggcgcggtgctcgaccagctcgacggcgcgggcgacgtgctggcgcgggaggtcgacggctgccaggacaacccgatcacctacgagggcgagctgctgcacggcggcaacttccacgccatgccggtgggtttcgcctccgaccagatcgggttggccatgcacatggccgcctacctggccgagcgccagctgggtctgctggtcagcccggtgaccaacggcgacctgccgcccatgctcaccccgcgcgccgggcgcggtgccgggctggccggggtgcagatcagcgcgacctcgttcgtctcgcggatccggcagctggtgttccccgcctcgctgaccaccctgccgaccaacggctggaaccaggaccacgtgccgatggcgctcaacggggcgaactcggtgttcgaggcgttggagctcggctggctgacggtcgggtcgctggcggtgggcgtcgcgcagctcgcggccatgaccggccacgccgcggagggcgtctgggcggagctggccgggatctgcccgccgctggacgccgaccgcccgctgggcgccgaggtgcgcgccgcgcgcgacctgctgtccgcgcacgcggaccaactgctcgtcgacgaggcagacgggaaggatttcggataataa
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