LOCUS       Exported                7401 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 7401)
  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 7401)
  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..7401
                     /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"
     RBS             629..651
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             659..1030
                     /codon_start=1
                     /product="Nitab4.5_0020038g0010 (solgenomics)"
                     /label=Nt_rbcS-T5
                     /translation="MQVWPPINKKKYETLSYLPDLSVEQLLREVEYLLKNGWVPCLEFE
                     TEHGFVYRENNKSPGYYDGRYWTMWKLPMFGCTDATQVLAEVEEAKKAYPQAWIRIIGF
                     DNVRQVQCISFIAYKPEGY"
     promoter        1113..1131
                     /label=T7P
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1132..1156
                     /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             1199..2380
                     /codon_start=1
                     /label=Nt-raf1
                     /note="Nt_Raf1"
                     /translation="MQQLYQPFRPPPPPLPPKFRNLDTNAKLEVLSNRLGLWYEYAPLI
                     PYLTSEGFTPSTLEEITGLTGVEQNRLVVAAQVRDTLVESAALDEETLSYFESGGAELL
                     YEIRLLSARQRTDAATFLVKNGFDAKQAQDLARAIKDYPRRRVDYGWDKFNGDSPGDCL
                     AFMYFRLAQEYAAAASEDLRRSSMEKALEVVESESARNLLVMELEGREVAKESVLDDGV
                     TVPLVRMKLGEVAESTIVVVLPVCKAEGRDVEVEAAPWECGGVGDFGIVEAEKDWRRWV
                     VLPGWQPIAGLERGGVAVSFKSGNFLPWREKSKYKQEPVLVVADRGRTEVASEDGFYLV
                     VDGGDGSNEEGLKVERGSTLKKRGVEQSLGIVLIVVRPPRWEDEEQLGEEDWD"
     promoter        2509..2527
                     /label=T7P
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2528..2552
                     /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             2567..2589
     CDS             2600..3115
                     /codon_start=1
                     /label=At-raf2
                     /note="RAF2"
                     /translation="MSNLAQDFLGDFGARDPYPEEIASQFGDKVLGCQSTEHKILIPNA
                     SVLSLSQLQCSPVSSSQPPLSGDDARTLLHKVLGWSIVDNEAGGLKIRCMWKVRDFGCG
                     VELINRIHKVAEASGHYPSLHLESPTQVRAELFTSSIGGLSMNDFIMAAKIDDIKTSDL
                     SPRKRAWA"
     terminator      3213..3260
                     /label=T7 term
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3427..3445
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3446..3470
                     /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             3485..3507
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3518..3760
                     /codon_start=1
                     /product="CV019724.1 without the transit peptide"
                     /label=Nt-BSDII
                     /translation="MATDSDKDTKVRSIVCQNCDGNGAVSCSQCKGSGVNSVDHFNGRF
                     KAGGLCWLCRGKKDVLCGDCNGAGFLGGFMSTFDE"
     terminator      3858..3905
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4362..5153)
                     /codon_start=1
                     /label=SmR
                     /note="SmR"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(5154..5245)
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
     rep_origin      complement(5293..6031)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="CloDF13 ori"
     CDS             complement(6241..7323)
                     /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(7324..7401)
                     /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 ttgccctcta gaaataattt tgtttaactt taagaaggag atatacatat
      661 gcaggtgtgg ccaccaatta acaagaagaa gtacgagact ctctcatacc ttcctgattt
      721 gagcgtggag caattgctta gggaagttga gtaccttttg aaaaatggat gggttccttg
      781 cttggaattc gagactgagc acggattcgt ctaccgtgag aacaacaagt caccaggtta
      841 ctacgatgga aggtactgga ccatgtggaa gttgcccatg ttcgggtgca ctgatgccac
      901 tcaggtcttg gctgaggtcg aggaggcaaa gaaggcttac ccacaagcct ggatcagaat
      961 cattggattc gacaacgtcc gtcaagtgca gtgcatcagt tttattgcct acaagcctga
     1021 aggctactaa tacttggcgc gcccagtgcg gccgcataat gcttaagtcg aacagaaagt
     1081 aatcgtattg tacacggccg cataatcgaa attaatacga ctcactatag gggaattgtg
     1141 agcggataac aattccccat cttagtatat tagttaagta taagaaggag atatacatat
     1201 gcaacaactt tatcagcctt ttagacctcc tcctccacct ctcccaccca aattccgcaa
     1261 tcttgacaca aatgcaaagc tcgaagttct atctaatcgc cttggtcttt ggtatgaata
     1321 tgctcctttg ataccttatt taacaagtga agggtttact ccttcaactc ttgaggaaat
     1381 cactggcctt actggagttg aacagaatcg gttggttgtt gctgcccaag ttcgtgatac
     1441 tcttgttgag tctgctgctt tagatgaaga aacgttgtcg tattttgaat ctggaggtgc
     1501 tgaattatta tatgaaattc gtttgcttag tgctaggcaa cgaactgatg cagcaacatt
     1561 tttagtgaag aatgggtttg atgcaaaaca agctcaagat ttagctagag caattaaaga
     1621 ttatccgcgt agacgtgtgg attatggttg ggacaagttt aatggtgatt ctcctggtga
     1681 ttgtttagct tttatgtact ttagattagc tcaagaatat gctgcggcag cttctgagga
     1741 tttgaggagg tcgtcaatgg agaaggcatt agaagttgtt gagtctgaaa gtgcgaggaa
     1801 tttgcttgtg atggagctag aaggacgtga ggttgctaag gagagtgtac ttgatgatgg
     1861 ggtgacagtg cctttggtga gaatgaagtt aggggaggta gctgagtcta cgattgtagt
     1921 cgttttgcct gtttgtaagg cggagggaag ggacgtggag gtggaggcgg cgccgtggga
     1981 atgtggtggc gtaggggatt ttggtatagt ggaggcggag aaggattgga ggagatgggt
     2041 tgttttgccc ggatggcaac ctatcgcggg gttagagaga ggtggagttg cagtgtcgtt
     2101 caagagcggg aactttttgc cgtggaggga gaagagtaag tacaagcaag agccggtttt
     2161 ggtagtggcg gataggggaa ggacagaagt ggcgtcggag gacgggtttt acttggtggt
     2221 tgatggtgga gacgggagca atgaggaggg attgaaagtg gaaagagggt caacattgaa
     2281 gaaaagaggg gttgaacaga gcttaggaat tgtgcttata gtagttaggc caccaaggtg
     2341 ggaagatgaa gaacaattag gtgaagaaga ttgggattaa agaccaagaa gcttagcttt
     2401 ctagactgca ggtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     2461 gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca
     2521 ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta
     2581 agaaggagat atacatatga tgtctaatct ggcgcaggat tttcttggtg acttcggagc
     2641 tcgtgaccct tacccggagg agatagcgag tcaattcgga gataaagtgt tgggatgcca
     2701 aagcactgag cacaagattt tgataccaaa cgcatctgtt ttgtctctct cccagcttca
     2761 gtgttcccct gtttcgtctt cacagcctcc tttgtccggc gatgatgcca gaactctcct
     2821 ccacaaggtt ttgggatgga gtatagtgga taatgaagcg ggtggtctga aaataaggtg
     2881 tatgtggaaa gtgagggatt ttgggtgcgg tgttgaactc ataaacagga tccataaggt
     2941 tgctgaagct tctggtcatt acccttctct tcacttggaa agtcctaccc aagttcgagc
     3001 tgaactattt acctcttcta tcggagggtt gagcatgaat gatttcataa tggcggctaa
     3061 aatagatgat atcaagactt ctgatctttc ccctaggaaa agagcttggg cgtgagctag
     3121 catgactggt ggacagcaaa tgggtcgcgg atccggctgc taacaaagcc cgaaaggaag
     3181 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     3241 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc
     3301 cggcagtacc ggcataacca agcactagtg tcggcgatat aggcgccagc aaccgcacct
     3361 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
     3421 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
     3481 taattttgtt taactttaag aaggagatat acatatgatg gccactgaca gtgataaaga
     3541 cacgaaagta aggagcatag tctgtcaaaa ttgtgatgga aatggtgcag tgtcgtgctc
     3601 acagtgcaaa ggctctgggg ttaactctgt agatcatttc aacgggcggt tcaaagctgg
     3661 tggactatgt tggttgtgca gaggtaaaaa agatgtgtta tgtggtgact gcaatggtgc
     3721 tggatttctt ggtggattta tgagtacgtt cgatgagtag gctagcatga ctggtggaca
     3781 gcaaatgggt cgcggatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     3841 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     3901 ttttgctgaa aggaggaact atatccggat atcccgcaag aggcccggca gtaccggcat
     3961 aaccaagcgg taccagcggt caattgtagt ggaggtgcta gcgcatgcgg aattggatat
     4021 cggccggcca cgcgatcgct gacgtcggta ccctcgagtc tggtaaagaa accgctgctg
     4081 cgaaatttga acgccagcac atggactcgt ctactagcgc agcttaatta acctaggctg
     4141 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcccttcct
     4201 caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag
     4261 caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcatcgtgg
     4321 ccggatcttg cggcccctcg gcttgaacga attgttagac attatttgcc gactaccttg
     4381 gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag
     4441 cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg
     4501 gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt
     4561 actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag
     4621 tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca
     4681 ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt
     4741 gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca
     4801 agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac
     4861 ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct
     4921 ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca
     4981 agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc
     5041 actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg
     5101 ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct catactcttc
     5161 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5221 gaatgtattt agaaaaataa acaaatagct agctcactcg gtcgctacgc tccgggcgtg
     5281 agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt ggataagcag
     5341 gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc cataggctcc
     5401 gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg ggagccgtga
     5461 ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag atccccaccg
     5521 tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt taccggatac
     5581 ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca cacactggta
     5641 tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac tccccgttca
     5701 gcccgactgc tgcgccttat ccggtaactg ttcacttgag tccaacccgg aaaagcacgg
     5761 taaaacgcca ctggcagcag ccattggtaa ctgggagttc gcagaggatt tgtttagcta
     5821 aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct acactggaag gacagatttg
     5881 gttgctgtgc tctgcgaaag ccagttacca cggttaagca gttccccaac tgacttaacc
     5941 ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga ttacgcgcag
     6001 aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct agatttcagt
     6061 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt
     6121 ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     6181 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     6241 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6301 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     6361 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     6421 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     6481 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
     6541 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     6601 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     6661 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     6721 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     6781 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     6841 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     6901 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     6961 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     7021 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     7081 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     7141 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     7201 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     7261 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     7321 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     7381 tttgcgccat tcgatggtgt c
//
