LOCUS Exported 7439 bp ds-DNA circular SYN 21-SEP-2020 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7439) AUTHORS Myat T. Lin, William D. Stone, Vishalsingh Chaudhari, Maureen Hanson TITLE Small subunits can determine enzyme kinetics of tobacco Rubisco expressed in Escherichia coli JOURNAL Nature Plants 2020, PMID: 32929197, DOI: 10.1038/s41477-020-00761-5 REFERENCE 2 (bases 1 to 7439) AUTHORS . TITLE Direct Submission JOURNAL Exported Monday, Sep 21, 2020 from SnapGene Viewer 5.0.8 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7439 /organism="recombinant plasmid" /mol_type="other DNA" promoter 46..64 /label=T7P /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 65..89 /label=lacO /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /note="The lac repressor binds to the lac operator toinhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." CDS 133..594 /codon_start=1 /product="XM_016647834 " /label=Nt-rbcX /translation="MAKAFKGNYKNLKKEKKSRRNLVVVSDLGGQYEYGFDDIKTQILN YFTFKAVRTILNQLNEMDSPQYQWFNDYVTVNKLRDGKRFIRNLAKEKQELAEKVMVTR LSLYAKWIKKCDHEEIYNRISDQNVEVMRERLMETVIWPSDDTNSERVG" promoter 617..635 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 667..689 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 697..1068 /codon_start=1 /product="Nitab4.5_0000682g0090 (solgenomics)" /label=Nt_rbcS-S2 /translation="MQVWPPINKKKYETLSYLPDLSQEQLLREVDYLLKNGWVPCLEFE TEHGFVYRENNKSPGYYDGRYWTMWKLPMFGCTDATQVLAEVEEAKKAYPQAWIRIIGF DNVRQVQCISFIAYKPEGY" promoter 1151..1169 /label=T7P /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1170..1194 /label=lacO /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /note="The lac repressor binds to the lac operator toinhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." CDS 1237..2418 /codon_start=1 /label=Nt-raf1 /note="Nt_Raf1" /translation="MQQLYQPFRPPPPPLPPKFRNLDTNAKLEVLSNRLGLWYEYAPLI PYLTSEGFTPSTLEEITGLTGVEQNRLVVAAQVRDTLVESAALDEETLSYFESGGAELL YEIRLLSARQRTDAATFLVKNGFDAKQAQDLARAIKDYPRRRVDYGWDKFNGDSPGDCL AFMYFRLAQEYAAAASEDLRRSSMEKALEVVESESARNLLVMELEGREVAKESVLDDGV TVPLVRMKLGEVAESTIVVVLPVCKAEGRDVEVEAAPWECGGVGDFGIVEAEKDWRRWV VLPGWQPIAGLERGGVAVSFKSGNFLPWREKSKYKQEPVLVVADRGRTEVASEDGFYLV VDGGDGSNEEGLKVERGSTLKKRGVEQSLGIVLIVVRPPRWEDEEQLGEEDWD" promoter 2547..2565 /label=T7P /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 2566..2590 /label=lacO /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /note="The lac repressor binds to the lac operator toinhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." RBS 2605..2627 CDS 2638..3153 /codon_start=1 /label=At-raf2 /note="RAF2" /translation="MSNLAQDFLGDFGARDPYPEEIASQFGDKVLGCQSTEHKILIPNA SVLSLSQLQCSPVSSSQPPLSGDDARTLLHKVLGWSIVDNEAGGLKIRCMWKVRDFGCG VELINRIHKVAEASGHYPSLHLESPTQVRAELFTSSIGGLSMNDFIMAAKIDDIKTSDL SPRKRAWA" terminator 3251..3298 /label=T7 term /note="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter 3465..3483 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 3484..3508 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." RBS 3523..3545 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 3556..3798 /codon_start=1 /product="CV019724.1 without the transit peptide" /label=Nt-BSDII /translation="MATDSDKDTKVRSIVCQNCDGNGAVSCSQCKGSGVNSVDHFNGRF KAGGLCWLCRGKKDVLCGDCNGAGFLGGFMSTFDE" terminator 3896..3943 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(4400..5191) /codon_start=1 /label=SmR /note="SmR" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" promoter complement(5192..5283) /gene="bla" /label=bla promoter /note="AmpR promoter" rep_origin complement(5331..6069) /direction=LEFT /label=CloDF13 ori /note="CloDF13 ori" CDS complement(6279..7361) /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="lacI" /note="The lac repressor binds to the lac operator toinhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(7362..7439) /gene="lacI" /label=lacI promoter /note="lacI promoter" ORIGIN 1 cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta 61 taggggaatt gtgagcggat aacaattccc ctgtagaaat aattttgttt aactttaata 121 aggagatata ccatggcaaa agcatttaag ggtaattaca agaatttaaa gaaggagaaa 181 aagagtagga ggaatcttgt ggttgtcagt gatttaggtg gacagtatga atatggcttt 241 gatgatatca aaacgcaaat tcttaactac ttcacattta aagcagtgag gactattttg 301 aaccagctta atgaaatgga ctctccacaa tatcaatggt tcaatgatta tgttacagta 361 aataagctta gggatgggaa gcgttttatt cgcaaccttg ctaaggagaa gcaagaactc 421 gctgaaaaag tgatggtaac acggctcagt ctgtatgcga aatggataaa aaaatgtgat 481 cacgaggaga tctacaaccg aatatctgat cagaatgtgg aagtgatgcg tgagcggctc 541 atggaaactg tgatttggcc atcagatgac acgaactcgg agagagttgg gtgaggcgcg 601 cggataacaa ttgccataat acgactcact atagggagac cacaacggtt tccctctaga 661 aataattttg tttaacttta agaaggagat atacatatgc aggtgtggcc accaattaac 721 aagaagaagt atgagacact ctcatacctt cctgatttga gccaggagca attgcttagg 781 gaagttgatt accttttgaa aaatggatgg gttccttgct tggaattcga gactgagcac 841 ggattcgtct accgtgagaa caataagtca ccagggtact acgatggtag atactggacc 901 atgtggaagt tgcccatgtt cgggtgcact gatgccactc aggtcttggc tgaggtagag 961 gaggcaaaga aggcttaccc acaggcctgg atcagaatca ttggattcga caacgtccgt 1021 caagtgcaat gcatcagttt catcgcctac aagcccgaag gctactaata cttggcgcgc 1081 ccagtgcggc cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta cacggccgca 1141 taatcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttccccatct 1201 tagtatatta gttaagtata agaaggagat atacatatgc aacaacttta tcagcctttt 1261 agacctcctc ctccacctct cccacccaaa ttccgcaatc ttgacacaaa tgcaaagctc 1321 gaagttctat ctaatcgcct tggtctttgg tatgaatatg ctcctttgat accttattta 1381 acaagtgaag ggtttactcc ttcaactctt gaggaaatca ctggccttac tggagttgaa 1441 cagaatcggt tggttgttgc tgcccaagtt cgtgatactc ttgttgagtc tgctgcttta 1501 gatgaagaaa cgttgtcgta ttttgaatct ggaggtgctg aattattata tgaaattcgt 1561 ttgcttagtg ctaggcaacg aactgatgca gcaacatttt tagtgaagaa tgggtttgat 1621 gcaaaacaag ctcaagattt agctagagca attaaagatt atccgcgtag acgtgtggat 1681 tatggttggg acaagtttaa tggtgattct cctggtgatt gtttagcttt tatgtacttt 1741 agattagctc aagaatatgc tgcggcagct tctgaggatt tgaggaggtc gtcaatggag 1801 aaggcattag aagttgttga gtctgaaagt gcgaggaatt tgcttgtgat ggagctagaa 1861 ggacgtgagg ttgctaagga gagtgtactt gatgatgggg tgacagtgcc tttggtgaga 1921 atgaagttag gggaggtagc tgagtctacg attgtagtcg ttttgcctgt ttgtaaggcg 1981 gagggaaggg acgtggaggt ggaggcggcg ccgtgggaat gtggtggcgt aggggatttt 2041 ggtatagtgg aggcggagaa ggattggagg agatgggttg ttttgcccgg atggcaacct 2101 atcgcggggt tagagagagg tggagttgca gtgtcgttca agagcgggaa ctttttgccg 2161 tggagggaga agagtaagta caagcaagag ccggttttgg tagtggcgga taggggaagg 2221 acagaagtgg cgtcggagga cgggttttac ttggtggttg atggtggaga cgggagcaat 2281 gaggagggat tgaaagtgga aagagggtca acattgaaga aaagaggggt tgaacagagc 2341 ttaggaattg tgcttatagt agttaggcca ccaaggtggg aagatgaaga acaattaggt 2401 gaagaagatt gggattaaag accaagaagc ttagctttct agactgcagg tcggcgatat 2461 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga 2521 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga 2581 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat acatatgatg 2641 tctaatctgg cgcaggattt tcttggtgac ttcggagctc gtgaccctta cccggaggag 2701 atagcgagtc aattcggaga taaagtgttg ggatgccaaa gcactgagca caagattttg 2761 ataccaaacg catctgtttt gtctctctcc cagcttcagt gttcccctgt ttcgtcttca 2821 cagcctcctt tgtccggcga tgatgccaga actctcctcc acaaggtttt gggatggagt 2881 atagtggata atgaagcggg tggtctgaaa ataaggtgta tgtggaaagt gagggatttt 2941 gggtgcggtg ttgaactcat aaacaggatc cataaggttg ctgaagcttc tggtcattac 3001 ccttctcttc acttggaaag tcctacccaa gttcgagctg aactatttac ctcttctatc 3061 ggagggttga gcatgaatga tttcataatg gcggctaaaa tagatgatat caagacttct 3121 gatctttccc ctaggaaaag agcttgggcg tgagctagca tgactggtgg acagcaaatg 3181 ggtcgcggat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc 3241 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct 3301 gaaaggagga actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag 3361 cactagtgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca 3421 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat 3481 aggggaattg tgagcggata acaattcccc tctagaaata attttgttta actttaagaa 3541 ggagatatac atatgatggc cactgacagt gataaagaca cgaaagtaag gagcatagtc 3601 tgtcaaaatt gtgatggaaa tggtgcagtg tcgtgctcac agtgcaaagg ctctggggtt 3661 aactctgtag atcatttcaa cgggcggttc aaagctggtg gactatgttg gttgtgcaga 3721 ggtaaaaaag atgtgttatg tggtgactgc aatggtgctg gatttcttgg tggatttatg 3781 agtacgttcg atgagtaggc tagcatgact ggtggacagc aaatgggtcg cggatccggc 3841 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc 3901 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat 3961 atccggatat cccgcaagag gcccggcagt accggcataa ccaagcggta ccagcggtca 4021 attgtagtgg aggtgctagc gcatgcggaa ttggatatcg gccggccacg cgatcgctga 4081 cgtcggtacc ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac gccagcacat 4141 ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact 4201 agcataaccc cttggggcct ctaaacgggt cccttcctca ggcatttgag aagcacacgg 4261 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt 4321 aacgaccctg ccctgaaccg acgaccgggt catcgtggcc ggatcttgcg gcccctcggc 4381 ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct ttcacgtagt 4441 ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct tgtccaagat 4501 aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc 4561 agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg 4621 acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg 4681 ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct 4741 ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca 4801 gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt 4861 ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa 4921 caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca 4981 aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca 5041 gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta 5101 cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag 5161 tcgatacttc ggcgatcacc gcttccctca tactcttcct ttttcaatat tattgaagca 5221 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 5281 aaatagctag ctcactcggt cgctacgctc cgggcgtgag actgcggcgg gcgctgcgga 5341 cacatacaaa gttacccaca gattccgtgg ataagcaggg gactaacatg tgaggcaaaa 5401 cagcagggcc gcgccggtgg cgtttttcca taggctccgc cctcctgcca gagttcacat 5461 aaacagacgc ttttccggtg catctgtggg agccgtgagg ctcaaccatg aatctgacag 5521 tacgggcgaa acccgacagg acttaaagat ccccaccgtt tccggcgggt cgctccctct 5581 tgcgctctcc tgttccgacc ctgccgttta ccggatacct gttccgcctt tctcccttac 5641 gggaagtgtg gcgctttctc atagctcaca cactggtatc tcggctcggt gtaggtcgtt 5701 cgctccaagc tgggctgtaa gcaagaactc cccgttcagc ccgactgctg cgccttatcc 5761 ggtaactgtt cacttgagtc caacccggaa aagcacggta aaacgccact ggcagcagcc 5821 attggtaact gggagttcgc agaggatttg tttagctaaa cacgcggttg ctcttgaagt 5881 gtgcgccaaa gtccggctac actggaagga cagatttggt tgctgtgctc tgcgaaagcc 5941 agttaccacg gttaagcagt tccccaactg acttaacctt cgatcaaacc acctccccag 6001 gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa aaaaaggatc tcaagaagat 6061 cctttgatct tttctactga accgctctag atttcagtgc aatttatctc ttcaaatgta 6121 gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc atgccccgcg 6181 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg 6241 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg 6301 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 6361 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc 6421 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg 6481 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg 6541 tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc 6601 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca 6661 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc 6721 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc 6781 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc 6841 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt 6901 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca 6961 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga 7021 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc 7081 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg 7141 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt 7201 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc 7261 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca 7321 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga 7381 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtc //