BBa_R0010 1 LacI promoter (lacI regulated) 2003-01-31T12:00:00Z 2015-05-08T01:14:14Z The Plac insert was PCR'd from the MG1655 strain of E.coli K12. Released HQ 2013 Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) <p> The pLac regulatory region is a 243 base-pair sequence with standard BioBrick prefix and suffix sections on its ends. It contains two protein binding sites: CAP, which is generally present in E.coli and is assocciated with cell health and availability of glucose., and LacI, the Lac inhibitor <bb_part>BBa_C0010</bb_part> which binds in an dimerized cooperative manner to inhibit the transcription of the protein that follows. In the presence of lactose or IPTG, an analog of lactose, LacI is unable to correctly bind and inhibit transcription. This allows <bb_part>BBa_R0010</bb_part> to be used as a inverter or as a detector of lactose or IPTG. false true _1_ 0 24 7 In stock false <P> <P><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 this regulator. This part is incompatible with environments containing lactose or lactose analogs. true annotation1961222 1 BBa_R0010 range1961222 1 1 200 annotation1961226 1 LacI binding site range1961226 1 166 200 annotation1961221 1 end of LacI coding region (inactive) range1961221 1 1 88 annotation1961223 1 CAP binding site range1961223 1 89 126 annotation1961224 1 -35 range1961224 1 137 142 annotation1961227 1 start range1961227 1 173 173 annotation1961225 1 -10 range1961225 1 161 166 BBa_K1088011 1 BBa_K1088011 B. subtilis dxs (lac promoter without lac inhibitor) 2013-08-27T11:00:00Z 2015-05-08T01:09:06Z The Lac promoter derives from E. coli. The Dxs protein is from B. sub This part has the B. sub dxs (BBa_K1088000) controlled by the common Lac promoter. Notice that there is no LacI in this system, which is required for regulation of the Lac promoter. false false _1398_ 0 12807 9 It's complicated false We forgot to acknowledge the need for high expression of LacI to keep the regulation of the lac promoter vaild. The constructs where assembled with USER cloning, and therefore there are no scarsites between the parts. There are however inserted scarsites between the RBS and the coding region aswell as between the lac promoter and the RBS to improve translational efficiency false Andreas Kj??r component2369133 1 BBa_K1088021 component2369138 1 BBa_K1088000 component2369136 1 BBa_J15001 component2369126 1 BBa_R0010 annotation2369133 1 BBa_K1088021 range2369133 1 201 206 annotation2369136 1 BBa_J15001 range2369136 1 207 216 annotation2369126 1 BBa_R0010 range2369126 1 1 200 annotation2369138 1 BBa_K1088000 range2369138 1 217 2118 BBa_K1088021 1 BBa_K1088021 Mixed site (ACTAGA) 2013-09-16T11:00:00Z 2015-05-08T01:09:06Z Restriction sites Mixed site (ACTAGA)when XbaI and SpeI sticky ends ligate false false _1398_ 0 17057 9 Not in stock false None false Patrick Rosendahl Andreassen 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 annotation1938045 1 SacI range1938045 1 1 3 annotation1938046 1 rbs range1938046 1 4 10 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_R0010_sequence 1 caatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacaca BBa_J15001_sequence 1 ctcaaggagg BBa_K1088011_sequence 1 caatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacaactagactcaaggaggatggatcttttatcaatacaggacccgtcgtttttaaaaaacatgtccattgatgaattagagaaattaagtgatgaaatccgtcagtttttaattacaagtttatccgcttccggcggccacatcggcccaaacttaggtgtcgtagagcttactgttgccctgcataaggaatttaacagcccgaaagacaaatttttatgggatgtaggccatcagtcgtatgtccataagctgctgacaggacgcggaaaagaatttgcgacgcttcgccagtacaaagggctttgcggatttccaaagcggagtgaaagcgagcacgatgtttgggaaaccgggcacagctcgacttctctgtcaggcgcgatgggaatggcagctgcccgtgatattaaaggaacggatgaatatattattccgatcattggtgacggcgcgctgaccggcggtatggcgctcgaagcccttaaccacatcggcgacgagaaaaaagacatgattgtcatccttaatgataatgaaatgagtattgcgccaaacgtcggtgccattcactctatgctcggacggctccgcactgcggggaaataccagtgggtcaaagatgagcttgaatacttatttaaaaagattccggcagttgggggcaagcttgccgccacggcggaacgggtcaaagacagcctgaaatacatgctcgtctccggaatgtttttcgaggagctcggttttacgtatttgggcccagtggacggacattcttatcatgagctgattgagaatcttcaatacgccaaaaaaacgaaaggccctgttcttctgcacgtcattacgaaaaaagggaaggggtacaaaccggctgagaccgatacgattgggacatggcatggtaccggaccatataaaattaataccggtgactttgtaaagccgaaagccgcagctccttcgtggagcggtcttgtcagcggaactgtgcagcgaatggcgcgcgaggacggacgcattgtagccattacgccggctatgcctgtcggttcaaagcttgaaggcttcgcaaaggaattccctgaccggatgttcgacgtaggaatcgcagaacagcatgccgcaacaatggctgcagctatggcaatgcagggtatgaagccgtttttggcgatttactcaaccttcctgcaaagggcatatgaccaagttgttcatgacatctgccgccaaaacgctaatgtgtttattggaattgaccgtgctggactcgttggcgctgatggagagacacatcaaggcgtgtttgatattgcgtttatgcgccacattccaaacatggtcttaatgatgccgaaagacgaaaatgaaggccagcacatggttcatacagcacttagctatgacgaaggcccgatagcaatgcgttttccgcgcggaaacggactcggcgtaaaaatggatgaacagttgaaaacgattccgatcggtacgtgggaggtgctgcgtccagggaacgatgctgtcatcttaacattcggcacaacaatcgaaatggcgattgaagcagccgaagagctgcagaaagaaggcctttccgtgcgcgttgtgaatgcgcgttttattaagccgattgatgaaaagatgatgaagagtatcctaaaagaaggcttgccaattttaacaattgaagaagcggtcttagaaggcggtttcggaagctcgattttagaattcgctcatgatcaaggtgaatatcatactccgattgacagaatgggtatacctgatcggtttattgaacacggaagtgtaacagcgcttcttgaggaaattggactgacaaaacagcaggtggcaaatcgtattagattactgatgccaccaaagacacacaaaggaattggatcatga BBa_K1088000_sequence 1 atggatcttttatcaatacaggacccgtcgtttttaaaaaacatgtccattgatgaattagagaaattaagtgatgaaatccgtcagtttttaattacaagtttatccgcttccggcggccacatcggcccaaacttaggtgtcgtagagcttactgttgccctgcataaggaatttaacagcccgaaagacaaatttttatgggatgtaggccatcagtcgtatgtccataagctgctgacaggacgcggaaaagaatttgcgacgcttcgccagtacaaagggctttgcggatttccaaagcggagtgaaagcgagcacgatgtttgggaaaccgggcacagctcgacttctctgtcaggcgcgatgggaatggcagctgcccgtgatattaaaggaacggatgaatatattattccgatcattggtgacggcgcgctgaccggcggtatggcgctcgaagcccttaaccacatcggcgacgagaaaaaagacatgattgtcatccttaatgataatgaaatgagtattgcgccaaacgtcggtgccattcactctatgctcggacggctccgcactgcggggaaataccagtgggtcaaagatgagcttgaatacttatttaaaaagattccggcagttgggggcaagcttgccgccacggcggaacgggtcaaagacagcctgaaatacatgctcgtctccggaatgtttttcgaggagctcggttttacgtatttgggcccagtggacggacattcttatcatgagctgattgagaatcttcaatacgccaaaaaaacgaaaggccctgttcttctgcacgtcattacgaaaaaagggaaggggtacaaaccggctgagaccgatacgattgggacatggcatggtaccggaccatataaaattaataccggtgactttgtaaagccgaaagccgcagctccttcgtggagcggtcttgtcagcggaactgtgcagcgaatggcgcgcgaggacggacgcattgtagccattacgccggctatgcctgtcggttcaaagcttgaaggcttcgcaaaggaattccctgaccggatgttcgacgtaggaatcgcagaacagcatgccgcaacaatggctgcagctatggcaatgcagggtatgaagccgtttttggcgatttactcaaccttcctgcaaagggcatatgaccaagttgttcatgacatctgccgccaaaacgctaatgtgtttattggaattgaccgtgctggactcgttggcgctgatggagagacacatcaaggcgtgtttgatattgcgtttatgcgccacattccaaacatggtcttaatgatgccgaaagacgaaaatgaaggccagcacatggttcatacagcacttagctatgacgaaggcccgatagcaatgcgttttccgcgcggaaacggactcggcgtaaaaatggatgaacagttgaaaacgattccgatcggtacgtgggaggtgctgcgtccagggaacgatgctgtcatcttaacattcggcacaacaatcgaaatggcgattgaagcagccgaagagctgcagaaagaaggcctttccgtgcgcgttgtgaatgcgcgttttattaagccgattgatgaaaagatgatgaagagtatcctaaaagaaggcttgccaattttaacaattgaagaagcggtcttagaaggcggtttcggaagctcgattttagaattcgctcatgatcaaggtgaatatcatactccgattgacagaatgggtatacctgatcggtttattgaacacggaagtgtaacagcgcttcttgaggaaattggactgacaaaacagcaggtggcaaatcgtattagattactgatgccaccaaagacacacaaaggaattggatcatga BBa_K1088021_sequence 1 actaga 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