BBa_K314110 1 f1 ori f1 origin 2010-10-07T11:00:00Z 2015-05-08T01:11:55Z Pet 29 vector Released HQ 2013 Phage origin recognized by M13. Used to make SS DNA when infected with M13k07 helper phage false false _496_ 0 6395 9 In stock true false IGEM 2010 Team Washington annotation2115768 1 f1 origin range2115768 1 18 324 BBa_K314103 1 BBa_K314103 Lac induced expression cassette 2010-10-07T11:00:00Z 2015-05-08T01:11:55Z construct Released HQ 2013 Lac induced protein expression insert includes f1 origin, Lac I gene and RBS false false _496_ 0 6395 9 In stock true false Tomas Huber, Justin Siegel component2091089 1 BBa_B0034 component2091082 1 BBa_K314111 component2091083 1 BBa_R0011 component2091081 1 BBa_K314110 annotation2091083 1 BBa_R0011 range2091083 1 1564 1617 annotation2091089 1 BBa_B0034 range2091089 1 1627 1638 annotation2091082 1 BBa_K314111 range2091082 1 464 1555 annotation2091081 1 BBa_K314110 range2091081 1 1 455 BBa_K314111 1 BBa_K314111 Lac I 2010-10-07T11:00:00Z 2015-05-08T01:11:55Z Pet 29 vector Lac I gene with promoter and RBS false false _496_ 0 6395 9 It's complicated false false IGEM 2010 Team Washington BBa_B0034 1 BBa_B0034 RBS (Elowitz 1999) -- defines RBS efficiency 2003-01-31T12:00:00Z 2015-08-31T04:07:20Z Released HQ 2013 RBS based on Elowitz repressilator. false true _1_ 0 24 7 In stock false Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. <p>No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info for this part: <a href="mailto:(bchow@media.mit.edu)">Brian Chow</a> true Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003. annotation23325 1 conserved range23325 1 5 8 BBa_K590034 1 FabH2 FabH2 2011-09-14T11:00:00Z 2015-05-08T01:12:48Z TBD Released HQ 2013 Makes Branched Chain Alkanes false false _761_ 0 10712 9 In stock true None false Matthew Harger BBa_R0011 1 lacI+pL Promoter (lacI regulated, lambda pL hybrid) 2003-01-31T12:00:00Z 2015-05-08T01:14:14Z represillator of Elowitz and Leibler (2000) Released HQ 2013 Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) <p> The PLlac 0-1 promoter is a hybrid regulatory region consisting of the promoter P(L) of phage lambda with the cI binding sites replaced with lacO1. The hybrid design allows for strong promotion that can nevertheless be tightly repressed by LacI, the Lac inhibitor (i.e. repressor) (<bb_part>BBa_C0010</bb_part>) ([LUTZ97]). The activity of the promoter can be regulated over a >600-fold range by IPTG in E.Coli DH5-alpha-Z1 (same paper reference). false true _1_ 0 24 7 In stock false <P> <P>hybrid promoter design to create strong promoter that is, at the same time, highly repressible. note that the upstream operator installed in this hybrid is slightly different than the one in the original source (Lutz and Bujard, 1997). the most upstream operator region is slightly truncated in the represillator version, so that both operators in the hybrid are the same sequence. see references for details. also, the sequence has been truncated after the transcriptional start site.<P>LacI binds to this regulator. This part is incompatible with species containing active LacI coding regions. Lactose and IPTG disable the operation of LacI and increase transcription. This part is incompatible with environments containing lactose or lactose analogs. true Neelaksh Varshney, Grace Kenney, Daniel Shen, Samantha Sutton annotation2001 1 lac O1 range2001 1 26 42 annotation7064 1 BBa_R0011 range7064 1 1 54 annotation1999 1 lac O1 range1999 1 3 19 annotation2000 1 -35 range2000 1 20 25 annotation2002 1 -10 range2002 1 43 48 BBa_K590064 1 BBa_K590064 The FabBrick: FabH2, an enzyme for changing up fatty acid biosynthesis 2011-10-24T11:00:00Z 2015-05-08T01:12:48Z Synthesized from the Bacillus subtilis protein sequence This part encodes FabH2. [http://2011.igem.org/Team:Washington 2011 University of Washington iGEM Team] has produced even chain length alkanes using this part and the [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590025 Petrobrick]. In addition, expression of this part and the [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590025 Petrobrick] should theoretically produce branched chain alkanes, but we have not been able to demonstrate this effect, possibly due to the absence of the appropriate substrates in ''E. coli'' ===Usage and Biology=== FabH2 is from ''Bacillus subtilis''. The FabH family of proteins initiates fatty acid elongation by converting an Acyl-CoA into an Acyl-ACP, with is extended by 2 carbon units to form longer chain length fatty acids. Normally, FabH proteins use a simple 2-carbon acetyl-CoA to start fatty acid biosynthesis, resulting in linear fatty acids. However, FabH2 can also use Isobutyryl-CoA, Isovaleryl-CoA, and 2-Methylbutyryl-CoA (products from Valine, Leucine, and Isoleucine degradation), resulting in 2-methyl branched fatty acid production. In addition, FabH2 has been hypothesized to start fatty acid elongation with a straight 3-carbon unit(propionyl-CoA), yielding odd chain length fatty acids, which could be converted into even chain length alkanes by the[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590025 Petrobrick]. Expression of FabH2 on the same high copy number constiuitive plasmid as the PetroBrick( as in Part [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590030 BBa_K590030]) results in slow cell growth( insert picture), and low alkane yield( under 10 mg/L vs. approximately 170 mg/L in the [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590025 Petrobrick]). In order to reduce these toxic effects, we cloned FabH2 onto a low copy number PSB1C3 [http://partsregistry.org/wiki/index.php?title=Part:BBa_K314103 IPTG inducible PetroBrick ]. This construct was co-transformed with the [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590025 Petrobrick] in XL1-Blue ''E. coli''. cells were grown up in rich TB media+ 5uM IPTG, and innocultated to OD10 in [http://2011.igem.org/Team:Washington/alkanebiosynthesis#100mL_M9_minGlucose_Media_Prep M9 production media] + 5uM IPTG. Alkanes were extracted after 24 hours. In addition, GC runs were performed on uninduced FabH2/Petrobrick cultures, and on cells expressing only the PetroBrick. [[Image:FabBrickGCMS.png|left|400px|thumb|GCMS trace confirming C16 alkane produced only upon FabBrick induction. ]] There was no significant difference between the GC peaks in the PetroBrick extract and in the uninduced FabH2/PetroBrick Extract. All of the peaks in these two extracts that fall within the expected elution time of the C16 alkane( 9.5-10 min) correspond to trace amounts of compounds that we cannot identify. When FabH2 is induced, we see a strong new peak at approximetly 9.75 minutes. The MS spectra of this peak is highly consistent with C16 alkane. The overall ion fragmentation fingerprint is identical to that alkane. Identification as a C16 alkane is confirmed by the presence of a strong parent ion at a mass of 226, exactly the mass of the C16 alkane. This confirms production of C16 alkane. In addition, some C14 alkane production was observed. This is the first time that even chain length alkanes have been produced recombinately, and expands the PetroBrick system to be able to produce all of the alkanes in the range C13-C17. Note that this peak corresponds to only trace levels of alkane( approximately 1mg/L), and future optimization is needed to increase even chain length production levels. false false _761_ 0 6386 9 It's complicated true Needed to dramatically lower expression of FabH2. false Matthew Harger component2160555 1 BBa_K590034 component2160554 1 BBa_K314103 annotation2160554 1 BBa_K314103 range2160554 1 1 1638 annotation2160555 1 BBa_K590034 range2160555 1 1645 2667 BBa_K314110_sequence 1 acgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaacaaaatattaacgtttacaatt BBa_B0034_sequence 1 aaagaggagaaa BBa_K314111_sequence 1 gtggtgaatgtgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggacatctcggtagtgggatacgacgataccgaagacagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtga BBa_K590034_sequence 1 atgtctaaagcgaaaatcaccgcgatcggcacctacgcgccgtctcgtcgcctcaccaacgcggacctcgaaaaaatcgtggacacgtctgacgagtggatcgttcagcgtaccggcatgcgtgaacgtcgtatcgcggacgaacaccagttcacctctgacctgtgcatcgaagcggttaaaaacctgaaatctcgttacaaaggtactctggacgatgttgacatgatcctggttgcgaccaccaccagcgactacgcgttcccgtctaccgcgtgccgcgttcaggagtacttcggttgggaatctaccggtgcgctcgacatcaacgctacgtgcgcaggcctcacttacggtctccacctggcgaatggcctgattacctctggcctgcaccagaagattctggttattgctggtgagactctctctaaagttaccgactataccgaccgtaccacctgtgttctcttcggtgacgccgcaggcgcgctgctcgtggagcgtgacgaggaaactccgggtttcctggcgtctgtgcagggtactagcggtaacggtggtgatatcctgtaccgtgcgggtctgcgtaacgaaatcaacggtgtacagctggttggttccggtaaaatggttcagaacggtcgtgaagtttacaaatgggccgctcgtaccgttccgggtgaatttgaacgcctgctgcacaaagcgggcctgtctagcgacgatctggactggtttgttccgcactctgcgaacctgcgtatgatcgaatctatctgcgagaaaaccccgttccctatcgagaagaccctgacctctgttgaacactacggtaacacctcttctgtttctatcgttctggcgctggatctggcggtgaaagcaggcaaactgaaaaaagaccaaattgttctgctgttcggtttcggcggtggtctgacttacaccggtctgctgatcaaatggggtatgggtggtagcggtggttctgactacaaagacgacgatgacaaataatga BBa_K590064_sequence 1 acgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaacaaaatattaacgtttacaatttactagaggtggtgaatgtgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggacatctcggtagtgggatacgacgataccgaagacagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgatactagagaattgtgagcggataacaattgacattgtgagcggataacaagatactgagcacatactagagaaagaggagaaatactagatgtctaaagcgaaaatcaccgcgatcggcacctacgcgccgtctcgtcgcctcaccaacgcggacctcgaaaaaatcgtggacacgtctgacgagtggatcgttcagcgtaccggcatgcgtgaacgtcgtatcgcggacgaacaccagttcacctctgacctgtgcatcgaagcggttaaaaacctgaaatctcgttacaaaggtactctggacgatgttgacatgatcctggttgcgaccaccaccagcgactacgcgttcccgtctaccgcgtgccgcgttcaggagtacttcggttgggaatctaccggtgcgctcgacatcaacgctacgtgcgcaggcctcacttacggtctccacctggcgaatggcctgattacctctggcctgcaccagaagattctggttattgctggtgagactctctctaaagttaccgactataccgaccgtaccacctgtgttctcttcggtgacgccgcaggcgcgctgctcgtggagcgtgacgaggaaactccgggtttcctggcgtctgtgcagggtactagcggtaacggtggtgatatcctgtaccgtgcgggtctgcgtaacgaaatcaacggtgtacagctggttggttccggtaaaatggttcagaacggtcgtgaagtttacaaatgggccgctcgtaccgttccgggtgaatttgaacgcctgctgcacaaagcgggcctgtctagcgacgatctggactggtttgttccgcactctgcgaacctgcgtatgatcgaatctatctgcgagaaaaccccgttccctatcgagaagaccctgacctctgttgaacactacggtaacacctcttctgtttctatcgttctggcgctggatctggcggtgaaagcaggcaaactgaaaaaagaccaaattgttctgctgttcggtttcggcggtggtctgacttacaccggtctgctgatcaaatggggtatgggtggtagcggtggttctgactacaaagacgacgatgacaaataatga BBa_R0011_sequence 1 aattgtgagcggataacaattgacattgtgagcggataacaagatactgagcaca BBa_K314103_sequence 1 acgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaacaaaatattaacgtttacaatttactagaggtggtgaatgtgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggacatctcggtagtgggatacgacgataccgaagacagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgatactagagaattgtgagcggataacaattgacattgtgagcggataacaagatactgagcacatactagagaaagaggagaaa 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