LOCUS       Exported                7428 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 7428)
  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 7428)
  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..7428
                     /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=T7P
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2552..2576
                     /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             2591..2613
                     /label=ParaR
     CDS             2624..3139
                     /codon_start=1
                     /label=At-raf2
                     /note="RAF2"
                     /translation="MSNLAQDFLGDFGARDPYPEEIASQFGDKVLGCQSTEHKILIPNA
                     SVLSLSQLQCSPVSSSQPPLSGDDARTLLHKVLGWSIVDNEAGGLKIRCMWKVRDFGCG
                     VELINRIHKVAEASGHYPSLHLESPTQVRAELFTSSIGGLSMNDFIMAAKIDDIKTSDL
                     SPRKRAWA"
     terminator      3237..3284
                     /label=T7 term
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3451..3469
                     /label=T7P
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3470..3494
                     /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             3509..3531
     CDS             3542..3787
                     /codon_start=1
                     /label=At-bsd2
                     /note="BSD2"
                     /translation="MAANNNPQGTKPNSLVCANCEGEGCVACSQCKGGGVNLIDHFNGQ
                     FKAGALCWLCRGKKEVLCGDCNGAGFIGGFLSTFDE"
     terminator      3885..3932
                     /label=T7 term
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4389..5180)
                     /codon_start=1
                     /label=SmR
                     /note="SmR"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(5181..5272)
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
     rep_origin      complement(5320..6058)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="CloDF13 ori"
     CDS             complement(6268..7350)
                     /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(7351..7428)
                     /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 atgatgtcta atctggcgca
     2641 ggattttctt ggtgacttcg gagctcgtga cccttacccg gaggagatag cgagtcaatt
     2701 cggagataaa gtgttgggat gccaaagcac tgagcacaag attttgatac caaacgcatc
     2761 tgttttgtct ctctcccagc ttcagtgttc ccctgtttcg tcttcacagc ctcctttgtc
     2821 cggcgatgat gccagaactc tcctccacaa ggttttggga tggagtatag tggataatga
     2881 agcgggtggt ctgaaaataa ggtgtatgtg gaaagtgagg gattttgggt gcggtgttga
     2941 actcataaac aggatccata aggttgctga agcttctggt cattaccctt ctcttcactt
     3001 ggaaagtcct acccaagttc gagctgaact atttacctct tctatcggag ggttgagcat
     3061 gaatgatttc ataatggcgg ctaaaataga tgatatcaag acttctgatc tttcccctag
     3121 gaaaagagct tgggcgtgag ctagcatgac tggtggacag caaatgggtc gcggatccgg
     3181 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     3241 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     3301 tatccggata tcccgcaaga ggcccggcag taccggcata accaagcact agtgtcggcg
     3361 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     3421 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
     3481 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat
     3541 gatggccgca aacaataatc ctcagggcac taaaccaaac agtttagttt gtgcaaattg
     3601 tgaaggggaa ggttgtgtgg catgttctca atgcaaagga ggtggtgtga atttgattga
     3661 tcatttcaat ggtcagttta aagctggtgc actctgttgg ctttgcagag gtaaaaagga
     3721 ggtgttgtgt ggggactgca atggagctgg tttcattggt gggttcttga gcaccttcga
     3781 tgagtaggct agcatgactg gtggacagca aatgggtcgc ggatccggct gctaacaaag
     3841 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     3901 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
     3961 ccgcaagagg cccggcagta ccggcataac caagcggtac cagcggtcaa ttgtagtgga
     4021 ggtgctagcg catgcggaat tggatatcgg ccggccacgc gatcgctgac gtcggtaccc
     4081 tcgagtctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta
     4141 ctagcgcagc ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc
     4201 ttggggcctc taaacgggtc ccttcctcag gcatttgaga agcacacggt cacactgctt
     4261 ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc
     4321 cctgaaccga cgaccgggtc atcgtggccg gatcttgcgg cccctcggct tgaacgaatt
     4381 gttagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct
     4441 tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta
     4501 gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg
     4561 acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc
     4621 actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca
     4681 tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga
     4741 cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg
     4801 atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc
     4861 agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact
     4921 tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg
     4981 atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata
     5041 tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac
     5101 gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg
     5161 gcgatcaccg cttccctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5221 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aatagctagc
     5281 tcactcggtc gctacgctcc gggcgtgaga ctgcggcggg cgctgcggac acatacaaag
     5341 ttacccacag attccgtgga taagcagggg actaacatgt gaggcaaaac agcagggccg
     5401 cgccggtggc gtttttccat aggctccgcc ctcctgccag agttcacata aacagacgct
     5461 tttccggtgc atctgtggga gccgtgaggc tcaaccatga atctgacagt acgggcgaaa
     5521 cccgacagga cttaaagatc cccaccgttt ccggcgggtc gctccctctt gcgctctcct
     5581 gttccgaccc tgccgtttac cggatacctg ttccgccttt ctcccttacg ggaagtgtgg
     5641 cgctttctca tagctcacac actggtatct cggctcggtg taggtcgttc gctccaagct
     5701 gggctgtaag caagaactcc ccgttcagcc cgactgctgc gccttatccg gtaactgttc
     5761 acttgagtcc aacccggaaa agcacggtaa aacgccactg gcagcagcca ttggtaactg
     5821 ggagttcgca gaggatttgt ttagctaaac acgcggttgc tcttgaagtg tgcgccaaag
     5881 tccggctaca ctggaaggac agatttggtt gctgtgctct gcgaaagcca gttaccacgg
     5941 ttaagcagtt ccccaactga cttaaccttc gatcaaacca cctccccagg tggttttttc
     6001 gtttacaggg caaaagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6061 ttctactgaa ccgctctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
     6121 cagccccata cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag
     6181 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     6241 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6301 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6361 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     6421 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     6481 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     6541 taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     6601 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     6661 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     6721 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     6781 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     6841 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     6901 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     6961 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     7021 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     7081 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     7141 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     7201 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     7261 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     7321 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     7381 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtc
//
