LOCUS       Exported                7422 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 7422)
  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 7422)
  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..7422
                     /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..1051
                     /codon_start=1
                     /product="trichome SSU from tobacco (XM_016595419)"
                     /label=Nt-RbcS-Tc
                     /translation="MKTWNPIDNKKFETLSYLPPLSEDSIAKEVDYMIQKGWIPCLEFD
                     QVGYVRRENSSMPGYYDGRYWTLWKLPMFGCNDSSQVLNEIQECKKAYPNAFIRCLAFD
                     NVKQVQCMAFLIQKPAAA"
     promoter        1134..1152
                     /label=T7P
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1153..1177
                     /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             1220..2401
                     /codon_start=1
                     /label=Nt-raf1
                     /note="Nt_Raf1"
                     /translation="MQQLYQPFRPPPPPLPPKFRNLDTNAKLEVLSNRLGLWYEYAPLI
                     PYLTSEGFTPSTLEEITGLTGVEQNRLVVAAQVRDTLVESAALDEETLSYFESGGAELL
                     YEIRLLSARQRTDAATFLVKNGFDAKQAQDLARAIKDYPRRRVDYGWDKFNGDSPGDCL
                     AFMYFRLAQEYAAAASEDLRRSSMEKALEVVESESARNLLVMELEGREVAKESVLDDGV
                     TVPLVRMKLGEVAESTIVVVLPVCKAEGRDVEVEAAPWECGGVGDFGIVEAEKDWRRWV
                     VLPGWQPIAGLERGGVAVSFKSGNFLPWREKSKYKQEPVLVVADRGRTEVASEDGFYLV
                     VDGGDGSNEEGLKVERGSTLKKRGVEQSLGIVLIVVRPPRWEDEEQLGEEDWD"
     promoter        2530..2548
                     /label=T7P
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2549..2573
                     /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             2588..2610
     CDS             2621..3136
                     /codon_start=1
                     /label=At-raf2
                     /note="RAF2"
                     /translation="MSNLAQDFLGDFGARDPYPEEIASQFGDKVLGCQSTEHKILIPNA
                     SVLSLSQLQCSPVSSSQPPLSGDDARTLLHKVLGWSIVDNEAGGLKIRCMWKVRDFGCG
                     VELINRIHKVAEASGHYPSLHLESPTQVRAELFTSSIGGLSMNDFIMAAKIDDIKTSDL
                     SPRKRAWA"
     terminator      3234..3281
                     /label=T7 term
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3448..3466
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3467..3491
                     /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             3506..3528
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3539..3781
                     /codon_start=1
                     /product="CV019724.1 without the transit peptide"
                     /label=Nt-BSDII
                     /translation="MATDSDKDTKVRSIVCQNCDGNGAVSCSQCKGSGVNSVDHFNGRF
                     KAGGLCWLCRGKKDVLCGDCNGAGFLGGFMSTFDE"
     terminator      3879..3926
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4383..5174)
                     /codon_start=1
                     /label=SmR
                     /note="SmR"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(5175..5266)
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
     rep_origin      complement(5314..6052)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="CloDF13 ori"
     CDS             complement(6262..7344)
                     /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(7345..7422)
                     /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 atatgaagac ttggaatccg attgacaaca agaaatttga
      721 gacactctca tatctgccac ctctatcaga agattcgatt gcaaaggagg ttgattacat
      781 gatccaaaag ggttggattc cttgccttga atttgatcag gtgggatatg tacgcaggga
      841 aaatagcagc atgccaggct actatgatgg aaggtattgg acactgtgga agctgcccat
      901 gtttggctgc aatgactcat cccaagttct caatgaaatc caagaatgca agaaagcgta
      961 cccaaatgct ttcattcgtt gtttggcatt tgacaatgtt aaacaggttc agtgcatggc
     1021 ctttctcatt cagaaacctg cagcagcata gtacttggcg cgcccagtgc ggccgcataa
     1081 tgcttaagtc gaacagaaag taatcgtatt gtacacggcc gcataatcga aattaatacg
     1141 actcactata ggggaattgt gagcggataa caattcccca tcttagtata ttagttaagt
     1201 ataagaagga gatatacata tgcaacaact ttatcagcct tttagacctc ctcctccacc
     1261 tctcccaccc aaattccgca atcttgacac aaatgcaaag ctcgaagttc tatctaatcg
     1321 ccttggtctt tggtatgaat atgctccttt gataccttat ttaacaagtg aagggtttac
     1381 tccttcaact cttgaggaaa tcactggcct tactggagtt gaacagaatc ggttggttgt
     1441 tgctgcccaa gttcgtgata ctcttgttga gtctgctgct ttagatgaag aaacgttgtc
     1501 gtattttgaa tctggaggtg ctgaattatt atatgaaatt cgtttgctta gtgctaggca
     1561 acgaactgat gcagcaacat ttttagtgaa gaatgggttt gatgcaaaac aagctcaaga
     1621 tttagctaga gcaattaaag attatccgcg tagacgtgtg gattatggtt gggacaagtt
     1681 taatggtgat tctcctggtg attgtttagc ttttatgtac tttagattag ctcaagaata
     1741 tgctgcggca gcttctgagg atttgaggag gtcgtcaatg gagaaggcat tagaagttgt
     1801 tgagtctgaa agtgcgagga atttgcttgt gatggagcta gaaggacgtg aggttgctaa
     1861 ggagagtgta cttgatgatg gggtgacagt gcctttggtg agaatgaagt taggggaggt
     1921 agctgagtct acgattgtag tcgttttgcc tgtttgtaag gcggagggaa gggacgtgga
     1981 ggtggaggcg gcgccgtggg aatgtggtgg cgtaggggat tttggtatag tggaggcgga
     2041 gaaggattgg aggagatggg ttgttttgcc cggatggcaa cctatcgcgg ggttagagag
     2101 aggtggagtt gcagtgtcgt tcaagagcgg gaactttttg ccgtggaggg agaagagtaa
     2161 gtacaagcaa gagccggttt tggtagtggc ggatagggga aggacagaag tggcgtcgga
     2221 ggacgggttt tacttggtgg ttgatggtgg agacgggagc aatgaggagg gattgaaagt
     2281 ggaaagaggg tcaacattga agaaaagagg ggttgaacag agcttaggaa ttgtgcttat
     2341 agtagttagg ccaccaaggt gggaagatga agaacaatta ggtgaagaag attgggatta
     2401 aagaccaaga agcttagctt tctagactgc aggtcggcga tataggcgcc agcaaccgca
     2461 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     2521 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     2581 aaataatttt gtttaacttt aagaaggaga tatacatatg atgtctaatc tggcgcagga
     2641 ttttcttggt gacttcggag ctcgtgaccc ttacccggag gagatagcga gtcaattcgg
     2701 agataaagtg ttgggatgcc aaagcactga gcacaagatt ttgataccaa acgcatctgt
     2761 tttgtctctc tcccagcttc agtgttcccc tgtttcgtct tcacagcctc ctttgtccgg
     2821 cgatgatgcc agaactctcc tccacaaggt tttgggatgg agtatagtgg ataatgaagc
     2881 gggtggtctg aaaataaggt gtatgtggaa agtgagggat tttgggtgcg gtgttgaact
     2941 cataaacagg atccataagg ttgctgaagc ttctggtcat tacccttctc ttcacttgga
     3001 aagtcctacc caagttcgag ctgaactatt tacctcttct atcggagggt tgagcatgaa
     3061 tgatttcata atggcggcta aaatagatga tatcaagact tctgatcttt cccctaggaa
     3121 aagagcttgg gcgtgagcta gcatgactgg tggacagcaa atgggtcgcg gatccggctg
     3181 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     3241 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     3301 ccggatatcc cgcaagaggc ccggcagtac cggcataacc aagcactagt gtcggcgata
     3361 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     3421 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
     3481 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatgat
     3541 ggccactgac agtgataaag acacgaaagt aaggagcata gtctgtcaaa attgtgatgg
     3601 aaatggtgca gtgtcgtgct cacagtgcaa aggctctggg gttaactctg tagatcattt
     3661 caacgggcgg ttcaaagctg gtggactatg ttggttgtgc agaggtaaaa aagatgtgtt
     3721 atgtggtgac tgcaatggtg ctggatttct tggtggattt atgagtacgt tcgatgagta
     3781 ggctagcatg actggtggac agcaaatggg tcgcggatcc ggctgctaac aaagcccgaa
     3841 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     3901 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tatcccgcaa
     3961 gaggcccggc agtaccggca taaccaagcg gtaccagcgg tcaattgtag tggaggtgct
     4021 agcgcatgcg gaattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt
     4081 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
     4141 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     4201 cctctaaacg ggtcccttcc tcaggcattt gagaagcaca cggtcacact gcttccggta
     4261 gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa
     4321 ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc ggcttgaacg aattgttaga
     4381 cattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa ttcttccaac
     4441 tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca
     4501 agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc
     4561 ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca
     4621 tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc
     4681 gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc tggacctacc
     4741 aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg
     4801 gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg
     4861 cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt gacttctaca
     4921 gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa
     4981 gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc aatatcactg
     5041 tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag caacgtcggt
     5101 tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac ttcggcgatc
     5161 accgcttccc tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5221 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagc tagctcactc
     5281 ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc ggacacatac aaagttaccc
     5341 acagattccg tggataagca ggggactaac atgtgaggca aaacagcagg gccgcgccgg
     5401 tggcgttttt ccataggctc cgccctcctg ccagagttca cataaacaga cgcttttccg
     5461 gtgcatctgt gggagccgtg aggctcaacc atgaatctga cagtacgggc gaaacccgac
     5521 aggacttaaa gatccccacc gtttccggcg ggtcgctccc tcttgcgctc tcctgttccg
     5581 accctgccgt ttaccggata cctgttccgc ctttctccct tacgggaagt gtggcgcttt
     5641 ctcatagctc acacactggt atctcggctc ggtgtaggtc gttcgctcca agctgggctg
     5701 taagcaagaa ctccccgttc agcccgactg ctgcgcctta tccggtaact gttcacttga
     5761 gtccaacccg gaaaagcacg gtaaaacgcc actggcagca gccattggta actgggagtt
     5821 cgcagaggat ttgtttagct aaacacgcgg ttgctcttga agtgtgcgcc aaagtccggc
     5881 tacactggaa ggacagattt ggttgctgtg ctctgcgaaa gccagttacc acggttaagc
     5941 agttccccaa ctgacttaac cttcgatcaa accacctccc caggtggttt tttcgtttac
     6001 agggcaaaag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6061 tgaaccgctc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
     6121 catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg
     6181 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     6241 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6301 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     6361 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     6421 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     6481 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     6541 gatgtccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     6601 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     6661 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     6721 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     6781 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     6841 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     6901 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     6961 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     7021 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     7081 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     7141 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     7201 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     7261 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     7321 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     7381 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tc
//
