LOCUS       Exported                7425 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 7425)
  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 7425)
  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..7425
                     /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
     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=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3470..3494
                     /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             3509..3531
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3542..3784
                     /codon_start=1
                     /product="CV019724.1 without the transit peptide"
                     /label=Nt-BSDII
                     /translation="MATDSDKDTKVRSIVCQNCDGNGAVSCSQCKGSGVNSVDHFNGRF
                     KAGGLCWLCRGKKDVLCGDCNGAGFLGGFMSTFDE"
     terminator      3882..3929
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4386..5177)
                     /codon_start=1
                     /label=SmR
                     /note="SmR"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(5178..5269)
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
     rep_origin      complement(5317..6055)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="CloDF13 ori"
     CDS             complement(6265..7347)
                     /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(7348..7425)
                     /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 gatggccact gacagtgata aagacacgaa agtaaggagc atagtctgtc aaaattgtga
     3601 tggaaatggt gcagtgtcgt gctcacagtg caaaggctct ggggttaact ctgtagatca
     3661 tttcaacggg cggttcaaag ctggtggact atgttggttg tgcagaggta aaaaagatgt
     3721 gttatgtggt gactgcaatg gtgctggatt tcttggtgga tttatgagta cgttcgatga
     3781 gtaggctagc atgactggtg gacagcaaat gggtcgcgga tccggctgct aacaaagccc
     3841 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     3901 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatcccg
     3961 caagaggccc ggcagtaccg gcataaccaa gcggtaccag cggtcaattg tagtggaggt
     4021 gctagcgcat gcggaattgg atatcggccg gccacgcgat cgctgacgtc ggtaccctcg
     4081 agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta
     4141 gcgcagctta attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg
     4201 gggcctctaa acgggtccct tcctcaggca tttgagaagc acacggtcac actgcttccg
     4261 gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct
     4321 gaaccgacga ccgggtcatc gtggccggat cttgcggccc ctcggcttga acgaattgtt
     4381 agacattatt tgccgactac cttggtgatc tcgcctttca cgtagtggac aaattcttcc
     4441 aactgatctg cgcgcgaggc caagcgatct tcttcttgtc caagataagc ctgtctagct
     4501 tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca
     4561 tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact
     4621 acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt
     4681 agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct
     4741 accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc
     4801 gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt
     4861 tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct
     4921 acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc
     4981 aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca
     5041 ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc
     5101 ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg
     5161 atcaccgctt ccctcatact cttccttttt caatattatt gaagcattta tcagggttat
     5221 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat agctagctca
     5281 ctcggtcgct acgctccggg cgtgagactg cggcgggcgc tgcggacaca tacaaagtta
     5341 cccacagatt ccgtggataa gcaggggact aacatgtgag gcaaaacagc agggccgcgc
     5401 cggtggcgtt tttccatagg ctccgccctc ctgccagagt tcacataaac agacgctttt
     5461 ccggtgcatc tgtgggagcc gtgaggctca accatgaatc tgacagtacg ggcgaaaccc
     5521 gacaggactt aaagatcccc accgtttccg gcgggtcgct ccctcttgcg ctctcctgtt
     5581 ccgaccctgc cgtttaccgg atacctgttc cgcctttctc ccttacggga agtgtggcgc
     5641 tttctcatag ctcacacact ggtatctcgg ctcggtgtag gtcgttcgct ccaagctggg
     5701 ctgtaagcaa gaactccccg ttcagcccga ctgctgcgcc ttatccggta actgttcact
     5761 tgagtccaac ccggaaaagc acggtaaaac gccactggca gcagccattg gtaactggga
     5821 gttcgcagag gatttgttta gctaaacacg cggttgctct tgaagtgtgc gccaaagtcc
     5881 ggctacactg gaaggacaga tttggttgct gtgctctgcg aaagccagtt accacggtta
     5941 agcagttccc caactgactt aaccttcgat caaaccacct ccccaggtgg ttttttcgtt
     6001 tacagggcaa aagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6061 tactgaaccg ctctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     6121 ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag
     6181 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     6241 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6301 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     6361 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     6421 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     6481 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     6541 cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     6601 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     6661 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     6721 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     6781 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     6841 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     6901 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     6961 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     7021 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     7081 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     7141 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     7201 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     7261 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     7321 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     7381 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtc
//
