BBa_K1796008 1 BBa_K1796008 nifD promoted from Paenibacillus sp. WLY78 2015-09-16T11:00:00Z 2015-09-17T08:19:32Z NifD(BBa_K1796008) is taken from genomic sequence of Paenibacillus sp.WLY78, we apply promotions on the sequences and synthesised it. Function: &#945;subunits of dinitrogease which also called FeMo protein. NifD encode a metallocluster: FeMo-co, a [Mo-7Fe-9S-C-homocitrate] cluster which serves as the active site of substrate binding and reduction. We abtained the sequences from genomes of Paenibacillus sp. WLY78. First, we promoted it ourselves&#65288;done by Nan Wang and Nannan Xie).Then we sent the sequences to synthesis, but unfortunately, striction enzyme cut site was involved after they promoted it again. But we were not informed of the error in promotion.After several times failed expriments, we found the problem.We tackled them over the matter, mistakes were corrected, but the correct genes can't arrive on time,it can only arrive after the deadline. Sequence submit is promoted by the synthesis company, containing a PstI in the gene. The sequence we promoted: CAGGAGGGAGGAATAGGCCAAATGTCTTCTATCGTTGACAAAGGTAAACAGATCGTTGAAGAAATCCTGGAAGTTTACCCGAAAAAAGCTAAAAAAGACCGTACCAAACACTTCGAAATCGCTGACGAAGAACTGGTTAACTGCGGTACCTGCTCTATCAAATCTAACATGAAATCTCGTCCGGGTGTTATGACCGCTCGTGGTTGCGCTTACGCTGGTTCTAAAGGTGTTGTTTGGGGTCCGATCAAAGACATGGTTCACATCTCTCACGGTCCGATCGGTTGCGGTCAGTACTCTTGGGGTACCCGTCGTAACTACGCTAACGGTATCCTGGGTATCGACAACTTCACCGCTATGCAGATCACCTCTAACTTCCAGGAAAAAGACATCGTTTTCGGTGGTGACAAAAAACTGGAAGTTATCTGCCGTGAAATCAAAGAAATGTTCCCGCTGGCTAAAGGTATCTCTGTTCAGTCTGAATGCCCGGTTGGTCTGATCGGTGACGACATCGGTGCTGTTGCTAAAAAAATGACCGAAGAACTGGGTATCCCGGTTATCCCGGTTCGTTGCGAAGGTTTCCGTGGTGTTTCTCAGTCTCTGGGTCACCACATCGCTAACGACGCTATCCGTGACTTCCTGATGGGTCGTCGTGAACTGAAAGAATGCGGTCCGTACGACGTTTCTATCATCGGTGACTACAACATCGGTGGTGACGCTTGGGCTTCTCGTATCCTGCTGGAAGAAATGGGTCTGCGTGTTATCGCTCAGTGGTCTGGTGACGGTACCATCAACGAACTGGGTATCGCTCACAAATCTAAACTGAACCTGATCCACTGCCACCGTTCTATGAACTACATGTGCACCACCATGGAACAGGAATACGGTATCCCGTGGATGGAATACAACTTCTTCGGTCCGACCAAAACCATGGAATCTCTGCGTGCTATCGCTGCTCGTTTCGACGAAACCATCCAGGAAAAATGCGAACAGGTTATCGCTCAGTACATGCCGCAGATGGAAGCTGTTATCCGTAAATACCGTCCGCGTCTGGAAGGTAAAAAAGTTATGCTGCTGATCGGTGGTCTGCGTGCTCGTCACACCATCGGTGCTTACGAAGACCTGGGTATGGAAATCGTTGCTACCGGTTACGAATTTGCTCACAAAGACGACTACGAAAAAACCTTCCCGGACGTTAAAGAAGGTACCATCCTGTACGACGACCCGACCGCTTACGAACTGGAAGAACTGGCTCAGCGTCTGAACATCGACCTGATGGGTGCTGGTGTTAAAGAAAAATACGTTTACCACAAAATGGGTATCCCGTTCCGTCAGATGCACTCTTGGGACTACTCTGGTCCATACCACGGGTTTGACGGCTTCAAGATCTTCGCGCGTGACATGGACATGACCATCAACTCTCCGGTTTGGTCTCTGCTGCCGTCTCGTCAGACCGCTGAAGTTCCGGTTTAATAA false false _2221_ 23592 23592 9 false We abtained the sequences from genomes of Paenibacillus sp. WLY78. First, we promoted it ourselves&#65288;done by Nan Wang and Nannan Xie).Then we sent the sequences to synthesis, but unfortunately, striction enzyme cut site was involved after they promoted it again. But the part can be assembled by gibson assembly,that is what we did. false Nannan Xie annotation2466009 1 nifD coding sequence range2466009 1 22 1473 annotation2475048 1 unknown rbs range2475048 1 1 21 BBa_B0015 1 BBa_B0015 double terminator (B0010-B0012) 2003-07-16T11:00:00Z 2015-08-31T04:07:20Z Released HQ 2013 Double terminator consisting of BBa_B0010 and BBa_B0012 false true _1_ 0 24 7 In stock false true Reshma Shetty component1916612 1 BBa_B0012 component1916610 1 BBa_B0010 annotation1916610 1 BBa_B0010 range1916610 1 1 80 annotation1916612 1 BBa_B0012 range1916612 1 89 129 BBa_K1796009 1 BBa_K1796009 nifK promoted from Paenibacillus sp. WLY78 2015-09-16T11:00:00Z 2015-09-17T08:42:28Z NifK(BBa_K1796009) is taken from genomic sequence of Paenibacillus sp.WLY78, we apply promotions on the sequences and synthesised it. Function: &#946; subunits of dinitrogease which also called FeMo protein.NifK encode a metallocluster: the P-cluster, a [8Fe-7S] cluster which shuttles electrons to FeMo-co. We abtained the sequences from genomes of Paenibacillus sp. WLY78. First, we promoted it ourselves&#65288;done by Nan Wang and Nannan Xie).Then we sent the sequences to synthesis. false false _2221_ 23592 23592 9 false We abtained the sequences from genomes of Paenibacillus sp. WLY78. First, we promoted it ourselves&#65288;done by Nan Wang and Nannan Xie).Then we sent the sequences to synthesis,the part can be assembled by gibson assembly,that is what we did. false Nannan Xie annotation2473682 1 nifK coding sequence range2473682 1 22 1554 annotation2475079 1 uknown rbs range2475079 1 1 21 BBa_K1796001 1 Pnif promoter of Paenibacillus sp. WLY78 nif gene cluster 2015-09-16T11:00:00Z 2015-09-18T02:35:42Z Pnif is from Paenibacillus sp. WLY78 genome,located upstream of nifB,the first gene of nitrogen fixation cluster. The promoter of Paenibacillus sp. WLY78 nif gene cluster, We named it Pnif. Pnif is a &#963;70-dependent promoter located upstream of nifB. Pnif is in control of all the gene clusters of nitrogen fixation genome. Confirmation of nif promoter We standardized the nif promoter of Paenibacillus sp. WLY78, and linked it to the upstream of 3 reporter genes (RFP reporter: BBa_K1357010, YFP reporter: BBa_E0430;amilCP reporter: BBa_K1357009) to confirm the reliability of this promoter. The results are shown in Figure 2. A B C http://2015.igem.org/wiki/images/b/b2/Confirmation_of_Pnif_AB.png http://2015.igem.org/wiki/images/thumb/3/34/C-confirmation_of_Pnif.jpeg/622px-C-confirmation_of_Pnif.jpeg Fig.4. Confirmation of nif promoter of Paenibacillus sp. WLY78. (A) Pnif+RFP reporter; (B) Pnif+YFP reporter;(C)Pnif+amilCP reporter. The results confirmed the reliability of Pnif. The initiation of Pnif does not require inducement. Thus it is a effective constitutive promoter. false false _2221_ 12637 23592 9 false effeciency of the promoter in E.coli for ordinary parts false Nannan Xie annotation2473661 1 Pnif range2473661 1 1 100 annotation2473637 1 -35 range2473637 1 62 67 annotation2473660 1 -10 range2473660 1 85 90 BBa_K1796016 1 BBa_K1796016 dinitrogease 2015-09-17T11:00:00Z 2015-09-18T10:02:26Z Paenibacillus sp. promoted by ourselves &#945;/&#946; subunits of dinitrogease false false _2221_ 23592 23592 9 false &#945;/&#946; subunits of dinitrogease false Nannan Xie component2475892 1 BBa_K1796008 component2475895 1 BBa_K1796009 component2475889 1 BBa_K1796001 component2475902 1 BBa_B0015 annotation2475892 1 BBa_K1796008 range2475892 1 109 1581 annotation2475889 1 BBa_K1796001 range2475889 1 1 100 annotation2475895 1 BBa_K1796009 range2475895 1 1590 3143 annotation2475902 1 BBa_B0015 range2475902 1 3152 3280 BBa_B0010 1 BBa_B0010 T1 from E. coli rrnB 2003-11-19T12:00:00Z 2015-08-31T04:07:20Z Transcriptional terminator consisting of a 64 bp stem-loop. false false _1_ 0 24 7 In stock false true Randy Rettberg annotation7018 1 BBa_B0010 range7018 1 1 80 annotation4184 1 stem_loop range4184 1 12 55 BBa_B0012 1 BBa_B0012 TE from coliphageT7 2003-01-31T12:00:00Z 2015-08-31T04:07:20Z Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>. Released HQ 2013 Transcription terminator for the <i>E.coli</i> RNA polymerase. false false _1_ 0 24 7 In stock false <P> <P>Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG).<P> Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator. false Reshma Shetty annotation1686 1 T7 TE range1686 1 8 27 annotation1690 1 polya range1690 1 28 41 annotation1687 1 stop range1687 1 34 34 annotation7020 1 BBa_B0012 range7020 1 1 41 BBa_B0010_sequence 1 ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctc BBa_K1796008_sequence 1 caggagggaggaataggccaaatgagcagcatcgtcgacaaaggcaaacagatcgtggaagaaatcctggaagtctacccgaaaaaagcgaaaaaagaccgcaccaaacacttcgagatcgcggacgaagaactggttaactgcggtacctgcagcatcaaaagcaacatgaaaagtcgtccgggcgttatgacggcacgtggttgtgcatacgcaggttcaaaaggcgttgtttggggtccgatcaaagacatggtccatattagccacggtccgattggttgcggtcaatattcttggggtacccgtcgtaactacgcaaacggtatcctgggcatcgataacttcaccgcaatgcagatcaccagcaacttccaggaaaaagacatcgtcttcggcggcgacaaaaaactggaagtcatctgccgcgaaatcaaagagatgtttccgctggcgaaaggcattagcgttcaatctgagtgtccggttggtctgattggcgacgatattggcgcggtcgccaaaaagatgaccgaagaactgggcattccggttattccggttcgttgcgaaggttttcgcggggttagtcagagcctgggtcatcatattgcaaacgacgcaattcgcgattttctgatgggccgtcgcgaactgaaagagtgcggtccgtacgacgttagcatcattggcgattacaacattggcggcgatgcttgggcaagtcgtattctgctggaagaaatgggtctgcgcgttattgcacagtggtctggcgacggtaccattaacgaactgggtatcgcgcacaaaagcaaactgaacctgatccactgccatcgttccatgaactacatgtgcaccaccatggaacaggaatacggtatcccgtggatggaatacaacttcttcggtccgaccaaaaccatggaaagcctgcgcgcaattgcagcgcgttttgacgaaaccatccaggagaaatgcgagcaggttattgcccagtatatgccgcagatggaggcggtcattcgtaaatatcgcccgcgtctggagggcaaaaaagttatgctgctgattggcggtctgcgcgcacgtcataccattggcgcatacgaagatctgggcatggaaattgttgcgaccggttacgaatttgcgcacaaagacgactacgagaaaaccttcccggacgtcaaagagggcaccattctgtacgacgatccgaccgcatacgaactggaagaactggcacagcgtctgaacattgatctgatgggcgcaggcgtcaaagagaaatacgtctaccacaaaatgggcattccgttccgtcagatgcatagctgggattatagcggtccgtatcacggtttcgacggctttaaaatcttcgcccgcgatatggatatgaccatcaacagcccggtttggagtctgctgccgtctcgtcaaaccgcagaagttccggtttaataa BBa_K1796016_sequence 1 cgtaaaatttgacacatatgtgaattgaggataaatgtcagggatttcatggagaagtgaattgactgtatttgtccctgtctctaagatgtaattatattactagagcaggagggaggaataggccaaatgagcagcatcgtcgacaaaggcaaacagatcgtggaagaaatcctggaagtctacccgaaaaaagcgaaaaaagaccgcaccaaacacttcgagatcgcggacgaagaactggttaactgcggtacctgcagcatcaaaagcaacatgaaaagtcgtccgggcgttatgacggcacgtggttgtgcatacgcaggttcaaaaggcgttgtttggggtccgatcaaagacatggtccatattagccacggtccgattggttgcggtcaatattcttggggtacccgtcgtaactacgcaaacggtatcctgggcatcgataacttcaccgcaatgcagatcaccagcaacttccaggaaaaagacatcgtcttcggcggcgacaaaaaactggaagtcatctgccgcgaaatcaaagagatgtttccgctggcgaaaggcattagcgttcaatctgagtgtccggttggtctgattggcgacgatattggcgcggtcgccaaaaagatgaccgaagaactgggcattccggttattccggttcgttgcgaaggttttcgcggggttagtcagagcctgggtcatcatattgcaaacgacgcaattcgcgattttctgatgggccgtcgcgaactgaaagagtgcggtccgtacgacgttagcatcattggcgattacaacattggcggcgatgcttgggcaagtcgtattctgctggaagaaatgggtctgcgcgttattgcacagtggtctggcgacggtaccattaacgaactgggtatcgcgcacaaaagcaaactgaacctgatccactgccatcgttccatgaactacatgtgcaccaccatggaacaggaatacggtatcccgtggatggaatacaacttcttcggtccgaccaaaaccatggaaagcctgcgcgcaattgcagcgcgttttgacgaaaccatccaggagaaatgcgagcaggttattgcccagtatatgccgcagatggaggcggtcattcgtaaatatcgcccgcgtctggagggcaaaaaagttatgctgctgattggcggtctgcgcgcacgtcataccattggcgcatacgaagatctgggcatggaaattgttgcgaccggttacgaatttgcgcacaaagacgactacgagaaaaccttcccggacgtcaaagagggcaccattctgtacgacgatccgaccgcatacgaactggaagaactggcacagcgtctgaacattgatctgatgggcgcaggcgtcaaagagaaatacgtctaccacaaaatgggcattccgttccgtcagatgcatagctgggattatagcggtccgtatcacggtttcgacggctttaaaatcttcgcccgcgatatggatatgaccatcaacagcccggtttggagtctgctgccgtctcgtcaaaccgcagaagttccggtttaataatactagaggcagactgcggaggtgccggtatgagcgaacgtccgaacatcgttgatcacaaccagctgttccgccaggataaatacgtccgtcaacgcgaagagaaacgcgcttttgaagcaccgtgtagtccggaagaagttaccgataccctggagtacaccaaaaccaaagagtacaaagacaaaaactttgcgcgcaccgcggttgttgttaatccggcaaaagcctgtcaaccgttaggcgcagttatggcagcactgggttttgagaaaaccctgccgtttattcacggctctcagggttgtaccgcatattttcgcagccatctggcacgtcatttcaaagaaccggttccggctgtttctaccagtatgacggaagacgcagctgtttttggcggtatgcgtaacctgatcgacggcattgaaaactgcattgcgctgtaccagccggaaatgattgcagtttgcaccacctgcatggcagaagttattggcgacgatctgagcgcatttctggcaaacgcacgtcaagaaggcgttttaccggaagatatgccggttccgtttgcaaacaccccgagttttagcggtagccatatcaccggctacgacgcaatgttacgtagcgtcctggaaaccctgtacaacaaaagcggtcgtaccgcacaaccgggtcacgaattaaaactgaacgtcctgctgggctttgacggttataccggtaacttcgcggaaatgcgtcgtatgctgggtatgtttggggcaacctataccatcctgggcgatcacagcagcaactttgattctggcgcgaccggcgaatatagctattattacggtggtaccccgttagaagacgttccgaaagcagcagacgcagcaggtaccttagcaattcagcagtacagcctgcgtaaaaccctgggctatatgaaacagacctggggtcagcaagttagcagcatttctaccccgctgggtattcgcgcaaccgatcgcttactggaagaaattagccgtttaagcggtcgcgaaattccggaagcactgaaacaggaacgcgcacgtattgttgacgctatgatggatagccacgcatatctgcacggtaaacgcgttgcaatggcaggcgatccggatatgttaatcggtctgatcggcttctgcttagaactgggcatggaaccggtacatatcgtttgcagcaacggcgatcgcaaattcgagaaagaagcggaactgctgctgaaatcctctccgtatggcgcagaagcaaccgttcattctggtcaggatctgtggcatatgcgtagcctgctgtttcaagatccggttgatctggctattggtagcagccatctgaaatttgcggcgaaagaggcggaaattccgttactgcgcgttggttttccgatctttgatcgccatcacctgcaccgttatccgattattggctatcagggcgcactgaatctgctgacccagtttgtcaacaccatcctggacgtcatggaagaacaagcaccggatcatagctttgacttagttcgttaataatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata BBa_K1796009_sequence 1 gcagactgcggaggtgccggtatgagcgaacgtccgaacatcgttgatcacaaccagctgttccgccaggataaatacgtccgtcaacgcgaagagaaacgcgcttttgaagcaccgtgtagtccggaagaagttaccgataccctggagtacaccaaaaccaaagagtacaaagacaaaaactttgcgcgcaccgcggttgttgttaatccggcaaaagcctgtcaaccgttaggcgcagttatggcagcactgggttttgagaaaaccctgccgtttattcacggctctcagggttgtaccgcatattttcgcagccatctggcacgtcatttcaaagaaccggttccggctgtttctaccagtatgacggaagacgcagctgtttttggcggtatgcgtaacctgatcgacggcattgaaaactgcattgcgctgtaccagccggaaatgattgcagtttgcaccacctgcatggcagaagttattggcgacgatctgagcgcatttctggcaaacgcacgtcaagaaggcgttttaccggaagatatgccggttccgtttgcaaacaccccgagttttagcggtagccatatcaccggctacgacgcaatgttacgtagcgtcctggaaaccctgtacaacaaaagcggtcgtaccgcacaaccgggtcacgaattaaaactgaacgtcctgctgggctttgacggttataccggtaacttcgcggaaatgcgtcgtatgctgggtatgtttggggcaacctataccatcctgggcgatcacagcagcaactttgattctggcgcgaccggcgaatatagctattattacggtggtaccccgttagaagacgttccgaaagcagcagacgcagcaggtaccttagcaattcagcagtacagcctgcgtaaaaccctgggctatatgaaacagacctggggtcagcaagttagcagcatttctaccccgctgggtattcgcgcaaccgatcgcttactggaagaaattagccgtttaagcggtcgcgaaattccggaagcactgaaacaggaacgcgcacgtattgttgacgctatgatggatagccacgcatatctgcacggtaaacgcgttgcaatggcaggcgatccggatatgttaatcggtctgatcggcttctgcttagaactgggcatggaaccggtacatatcgtttgcagcaacggcgatcgcaaattcgagaaagaagcggaactgctgctgaaatcctctccgtatggcgcagaagcaaccgttcattctggtcaggatctgtggcatatgcgtagcctgctgtttcaagatccggttgatctggctattggtagcagccatctgaaatttgcggcgaaagaggcggaaattccgttactgcgcgttggttttccgatctttgatcgccatcacctgcaccgttatccgattattggctatcagggcgcactgaatctgctgacccagtttgtcaacaccatcctggacgtcatggaagaacaagcaccggatcatagctttgacttagttcgttaataa BBa_K1796001_sequence 1 cgtaaaatttgacacatatgtgaattgaggataaatgtcagggatttcatggagaagtgaattgactgtatttgtccctgtctctaagatgtaattatat BBa_B0012_sequence 1 tcacactggctcaccttcgggtgggcctttctgcgtttata BBa_B0015_sequence 1 ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata 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