LOCUS       Exported                7404 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 7404)
  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 7404)
  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..7404
                     /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        603..621
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             653..675
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             683..1054
                     /codon_start=1
                     /product="Nt-rbcS-S1 (XM_016626460)"
                     /label=Nt-rbcS-S1
                     /translation="MQVWPPINKKKYETLSYLPDLSQEQLLSEVEYLLKNGWVPCLEFE
                     TEHGFVYRENNKSPGYYDGRYWTMWKLPMFGCTDATQVLAEVEEAKKAYPQAWIRIIGF
                     DNVRQVQCISFIAYKPEGY"
     promoter        1137..1155
                     /label=T7P
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1156..1180
                     /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             1223..2404
                     /codon_start=1
                     /label=Nt-raf1
                     /note="Nt_Raf1"
                     /translation="MQQLYQPFRPPPPPLPPKFRNLDTNAKLEVLSNRLGLWYEYAPLI
                     PYLTSEGFTPSTLEEITGLTGVEQNRLVVAAQVRDTLVESAALDEETLSYFESGGAELL
                     YEIRLLSARQRTDAATFLVKNGFDAKQAQDLARAIKDYPRRRVDYGWDKFNGDSPGDCL
                     AFMYFRLAQEYAAAASEDLRRSSMEKALEVVESESARNLLVMELEGREVAKESVLDDGV
                     TVPLVRMKLGEVAESTIVVVLPVCKAEGRDVEVEAAPWECGGVGDFGIVEAEKDWRRWV
                     VLPGWQPIAGLERGGVAVSFKSGNFLPWREKSKYKQEPVLVVADRGRTEVASEDGFYLV
                     VDGGDGSNEEGLKVERGSTLKKRGVEQSLGIVLIVVRPPRWEDEEQLGEEDWD"
     promoter        2533..2551
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2552..2576
                     /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             2591..2613
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2621..3118
                     /codon_start=1
                     /product="NW_015822561.1"
                     /label=Nt-raf2
                     /translation="MLLGDFGARDPFPAEIESKFGEKVLGNTDTEHKILIPAASALSLA
                     QQDCIVISPGQTPLSEYEARQLLFKVVGWRLAHEDGVLKLQCTWKLRDFECGVELINRI
                     GKVVEGTGHVPTLLHLEQSNQVRAELWTASIGGLSINDFIVAAKIDQIKTSDLVPRKRV
                     WA"
     terminator      3216..3263
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3430..3448
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3449..3473
                     /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             3488..3510
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3521..3763
                     /codon_start=1
                     /product="CV019724.1 without the transit peptide"
                     /label=Nt-BSDII
                     /translation="MATDSDKDTKVRSIVCQNCDGNGAVSCSQCKGSGVNSVDHFNGRF
                     KAGGLCWLCRGKKDVLCGDCNGAGFLGGFMSTFDE"
     terminator      3861..3908
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4365..5156)
                     /codon_start=1
                     /label=SmR
                     /note="SmR"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(5157..5248)
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
     rep_origin      complement(5296..6034)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="CloDF13 ori"
     CDS             complement(6244..7326)
                     /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(7327..7404)
                     /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 cgtaatacga ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta
      661 actttaagaa ggagatatac atatgcaggt gtggccacca attaacaaga agaagtacga
      721 gactctctca taccttcctg atttgagcca ggagcaattg cttagtgaag ttgagtacct
      781 tttgaaaaat ggatgggttc cttgcttgga attcgagact gagcacggat ttgtctatcg
      841 tgaaaacaac aagtcaccag gatactatga tggcagatac tggaccatgt ggaagctacc
      901 tatgttcgga tgcactgatg ccacccaagt gttggctgag gtagaagagg cgaagaaggc
      961 atacccacag gcctggatcc gtatcattgg attcgacaac gtgcgtcaag tgcagtgcat
     1021 cagtttcatt gcctacaagc ctgaaggata ttaatacttg gcgcgcccag tgcggccgca
     1081 taatgcttaa gtcgaacaga aagtaatcgt attgtacacg gccgcataat cgaaattaat
     1141 acgactcact ataggggaat tgtgagcgga taacaattcc ccatcttagt atattagtta
     1201 agtataagaa ggagatatac atatgcaaca actttatcag ccttttagac ctcctcctcc
     1261 acctctccca cccaaattcc gcaatcttga cacaaatgca aagctcgaag ttctatctaa
     1321 tcgccttggt ctttggtatg aatatgctcc tttgatacct tatttaacaa gtgaagggtt
     1381 tactccttca actcttgagg aaatcactgg ccttactgga gttgaacaga atcggttggt
     1441 tgttgctgcc caagttcgtg atactcttgt tgagtctgct gctttagatg aagaaacgtt
     1501 gtcgtatttt gaatctggag gtgctgaatt attatatgaa attcgtttgc ttagtgctag
     1561 gcaacgaact gatgcagcaa catttttagt gaagaatggg tttgatgcaa aacaagctca
     1621 agatttagct agagcaatta aagattatcc gcgtagacgt gtggattatg gttgggacaa
     1681 gtttaatggt gattctcctg gtgattgttt agcttttatg tactttagat tagctcaaga
     1741 atatgctgcg gcagcttctg aggatttgag gaggtcgtca atggagaagg cattagaagt
     1801 tgttgagtct gaaagtgcga ggaatttgct tgtgatggag ctagaaggac gtgaggttgc
     1861 taaggagagt gtacttgatg atggggtgac agtgcctttg gtgagaatga agttagggga
     1921 ggtagctgag tctacgattg tagtcgtttt gcctgtttgt aaggcggagg gaagggacgt
     1981 ggaggtggag gcggcgccgt gggaatgtgg tggcgtaggg gattttggta tagtggaggc
     2041 ggagaaggat tggaggagat gggttgtttt gcccggatgg caacctatcg cggggttaga
     2101 gagaggtgga gttgcagtgt cgttcaagag cgggaacttt ttgccgtgga gggagaagag
     2161 taagtacaag caagagccgg ttttggtagt ggcggatagg ggaaggacag aagtggcgtc
     2221 ggaggacggg ttttacttgg tggttgatgg tggagacggg agcaatgagg agggattgaa
     2281 agtggaaaga gggtcaacat tgaagaaaag aggggttgaa cagagcttag gaattgtgct
     2341 tatagtagtt aggccaccaa ggtgggaaga tgaagaacaa ttaggtgaag aagattggga
     2401 ttaaagacca agaagcttag ctttctagac tgcaggtcgg cgatataggc gccagcaacc
     2461 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
     2521 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
     2581 tagaaataat tttgtttaac tttaagaagg agatatacat atgctattgg gcgattttgg
     2641 ggcgagagat ccattcccag ctgaaataga gagtaaattc ggtgaaaaag ttttgggtaa
     2701 tactgacacg gagcacaaaa ttctcatccc agctgcttct gctctgtccc ttgctcagca
     2761 agactgcatt gtcatctcac ctggtcaaac tcctctctcg gaatatgaag ctcgacagct
     2821 cctattcaag gttgttggct ggagactggc acatgaagat ggggtgctca agctacaatg
     2881 tacatggaaa ttaagagact tcgaatgtgg ggttgaactt atcaatcgaa taggcaaagt
     2941 ggtggaaggc actgggcacg ttcccacact tcttcactta gagcagtcca atcaagttcg
     3001 tgctgagcta tggactgctt ccatcggagg tttgagcata aacgatttca ttgtagctgc
     3061 taaaatagat cagataaaga catcggacct tgtcccaaga aaaagagttt gggcataagc
     3121 tagcatgact ggtggacagc aaatgggtcg cggatccggc tgctaacaaa gcccgaaagg
     3181 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     3241 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatat cccgcaagag
     3301 gcccggcagt accggcataa ccaagcacta gtgtcggcga tataggcgcc agcaaccgca
     3361 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     3421 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     3481 aaataatttt gtttaacttt aagaaggaga tatacatatg atggccactg acagtgataa
     3541 agacacgaaa gtaaggagca tagtctgtca aaattgtgat ggaaatggtg cagtgtcgtg
     3601 ctcacagtgc aaaggctctg gggttaactc tgtagatcat ttcaacgggc ggttcaaagc
     3661 tggtggacta tgttggttgt gcagaggtaa aaaagatgtg ttatgtggtg actgcaatgg
     3721 tgctggattt cttggtggat ttatgagtac gttcgatgag taggctagca tgactggtgg
     3781 acagcaaatg ggtcgcggat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     3841 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     3901 gttttttgct gaaaggagga actatatccg gatatcccgc aagaggcccg gcagtaccgg
     3961 cataaccaag cggtaccagc ggtcaattgt agtggaggtg ctagcgcatg cggaattgga
     4021 tatcggccgg ccacgcgatc gctgacgtcg gtaccctcga gtctggtaaa gaaaccgctg
     4081 ctgcgaaatt tgaacgccag cacatggact cgtctactag cgcagcttaa ttaacctagg
     4141 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtccctt
     4201 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     4261 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcatcg
     4321 tggccggatc ttgcggcccc tcggcttgaa cgaattgtta gacattattt gccgactacc
     4381 ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc
     4441 aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac
     4501 tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg
     4561 gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc atcgccagcc
     4621 cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt
     4681 tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct
     4741 cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc gaagatacct
     4801 gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc
     4861 cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc
     4921 tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca
     4981 tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt caggccgcca
     5041 tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg
     5101 acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc cctcatactc
     5161 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5221 tttgaatgta tttagaaaaa taaacaaata gctagctcac tcggtcgcta cgctccgggc
     5281 gtgagactgc ggcgggcgct gcggacacat acaaagttac ccacagattc cgtggataag
     5341 caggggacta acatgtgagg caaaacagca gggccgcgcc ggtggcgttt ttccataggc
     5401 tccgccctcc tgccagagtt cacataaaca gacgcttttc cggtgcatct gtgggagccg
     5461 tgaggctcaa ccatgaatct gacagtacgg gcgaaacccg acaggactta aagatcccca
     5521 ccgtttccgg cgggtcgctc cctcttgcgc tctcctgttc cgaccctgcc gtttaccgga
     5581 tacctgttcc gcctttctcc cttacgggaa gtgtggcgct ttctcatagc tcacacactg
     5641 gtatctcggc tcggtgtagg tcgttcgctc caagctgggc tgtaagcaag aactccccgt
     5701 tcagcccgac tgctgcgcct tatccggtaa ctgttcactt gagtccaacc cggaaaagca
     5761 cggtaaaacg ccactggcag cagccattgg taactgggag ttcgcagagg atttgtttag
     5821 ctaaacacgc ggttgctctt gaagtgtgcg ccaaagtccg gctacactgg aaggacagat
     5881 ttggttgctg tgctctgcga aagccagtta ccacggttaa gcagttcccc aactgactta
     5941 accttcgatc aaaccacctc cccaggtggt tttttcgttt acagggcaaa agattacgcg
     6001 cagaaaaaaa ggatctcaag aagatccttt gatcttttct actgaaccgc tctagatttc
     6061 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
     6121 attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     6181 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     6241 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6301 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     6361 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     6421 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     6481 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
     6541 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     6601 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     6661 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     6721 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     6781 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     6841 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     6901 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     6961 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     7021 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     7081 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     7141 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     7201 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     7261 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     7321 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     7381 ggttttgcgc cattcgatgg tgtc
//
