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_0000912g0270 (solgenomics)"
                     /label=Nt_rbcS-T4a
                     /translation="MQVWPPINKKKYETLSYLPDLSEEQLLREVEYLLKNGWVPCLEFE
                     TEHGFVYRENNKSPGYYDGRYWTMRKLPMFGCTDATQVLAEVEEAKKAYPQAWIRIIGF
                     DNVRQVQCISFIAYKPAGY"
     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 acgagacact ctcatacctt cctgatttga gcgaggagca attgcttagg
      781 gaagttgaat accttttgaa aaatggatgg gttccttgct tggaattcga gactgagcac
      841 ggattcgtct accgtgagaa taacaagtca ccaggttact acgatggaag gtactggacc
      901 atgaggaagt tgcccatgtt cgggtgcact gatgccactc aggtcttggc tgaggtcgag
      961 gaggcaaaga aggcttaccc acaagcctgg atcagaatca ttggattcga caacgtccgt
     1021 caagtgcaat gcatcagttt catcgcctac aagcccgcag 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
//
