LOCUS       Exported                8937 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 8937)
  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 8937)
  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..8937
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        104..208
                     /note="AmpR promoter"
     CDS             209..1069
                     /codon_start=1
                     /label=AmpR
                     /note="AmpR"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1240..1828
                     /direction=RIGHT
                     /label=ori
                     /note="ori"
     CDS             complement(2258..2449)
                     /codon_start=1
                     /label=rop
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        3128..3146
                     /label=T7P
                     /note="T7 promoter"
     protein_bind    3147..3171
                     /label=lacO
                     /bound_moiety="LacI"
                     /note="lac operator"
     CDS             3216..4838
                     /codon_start=1
                     /product="XM_016653834"
                     /label=Nt_cpn60alpha
                     /translation="MAKEIAFDQKSRSALQAGIDKLADAVGLTLGPRGRNVVLDEYGTP
                     KVVNDGVTIARAIELPDAMENAGAALIREVASKTNDSAGDGTTTASVLAREIIKLGLLS
                     VTSGANPVSLKKGIDKTVLGLIEELEKKARPVKGRDDIKAIASISAGNDESIGTMIADA
                     IDKVGPDGVLSIESSSSFETTVHVEEGMEIDRGYISPQFVTNPEKLIVEFENARVLVTD
                     QKISAIKDIIPLLEKTTQLRSPLLIIAEDVTGEALATLVVNKLRGILNVAAIKAPGFGE
                     RRKALLQDIAIVTGAEYQATDLGLLIEGTPVEALGIARKVTITKDSTTIIADAASKDEI
                     QARIAQLKKELSETDSVYDSEKLAERIAKLSGGVAVIKVGAATEAELEDRKLRIEDAKN
                     ATFAAIEEGIVPGGGAAFVHLSTYVPAIKDKLEDADEKLGADIIQRALVAPASLIAQNA
                     GIEGEVVVEKVREAEWEMGYNAMTDKYENLVDAGVIDPAKVTRCALQNSASVAGMVLTT
                     QAIVVEKAKPKAAAPAAPQGLSV"
     promoter        4866..4884
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4916..4938
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4946..5239
                     /codon_start=1
                     /label=groES
                     /note="groES gene from E. coli"
                     /translation="MNIRPLHDRVIVKRKEVETKSAGGIVLTGSAAAKSTRGEVLAVGN
                     GRILENGEVKPLDVKVGDIVIFNDGYGVKSEKIDNEEVLIMSESDILAIVEA"
     RBS             5315..5337
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5349..6782
                     /codon_start=1
                     /product="Rubisco large subunit from N. tabacum"
                     /label=Nt-rbcL
                     /translation="MSPQTETKASVGFKAGVKEYKLTYYTPEYQTKDTDILAAFRVTPQ
                     PGVPPEEAGAAVAAESSTGTWTTVWTDGLTSLDRYKGRCYRIERVVGEKDQYIAYVAYP
                     LDLFEEGSVTNMFTSIVGNVFGFKALRALRLEDLRIPPAYVKTFQGPPHGIQVERDKLN
                     KYGRPLLGCTIKPKLGLSAKNYGRAVYECLRGGLDFTKDDENVNSQPFMRWRDRFLFCA
                     EALYKAQAETGEIKGHYLNATAGTCEEMIKRAVFARELGVPIVMHDYLTGGFTANTSLA
                     HYCRDNGLLLHIHRAMHAVIDRQKNHGIHFRVLAKALRMSGGDHIHSGTVVGKLEGERD
                     ITLGFVDLLRDDFVEQDRSRGIYFTQDWVSLPGVLPVASGGIHVWHMPALTEIFGDDSV
                     LQFGGGTLGHPWGNAPGAVANRVALEACVKARNEGRDLAQEGNEIIREACKWSPELAAA
                     CEVWKEIVFNFAAVDVLDK"
     promoter        6865..6883
                     /label=T7P
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6884..6908
                     /label=lacO
                     /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)."
     CDS             6957..8600
                     /codon_start=1
                     /product="99% identical to XM_016634979"
                     /label=Nt-cpn60-beta-2
                     /translation="MAAKELYFNKDGSAIKRLQVGVNKLADLVGVTLGPKGRNVVLESK
                     YGAPKIVNDGVTVAREVELEDPVENIGAKLVRQAAAKTNDLAGDGTTTSVVLAQGLVAE
                     GVKVVAAGANPVLITRGIEKTTKALVAELKNMSKEVEDSELADVAAVSAGNNSEVGSMI
                     AEAMSKVGRKGVVTLEEGKSAENSLRVVEGMQFDRGYISPYFVTDSEKMSVEYENCKLL
                     LVDKKITNARDLVGVLEDAIRNGYPILIIAEDIEQEALATLVVNKLRGALKVAALKAPG
                     FGERKSQYLDDIATLTGGTVIREELGLTLDKADKEVLGHASKVVLTKDATTIVGDGSTQ
                     EAVNKRVVQIKNLIEAADQDYEKEKLNERIAKLSGGVAVIQVGAQTETELKEKKLRVED
                     ALNATKAAVEEGIVVGGGCTLLRLASKVDAIKETLENDEEKVGADIVKRALSYPLKLIA
                     KNAGVNGSVVSEKVLSSDNPKFGYNAATGNYEDLMAAGIIDPTKVVRCCLEHASSVAKT
                     FLMSDCVVVEIKEPEPAVAGNPMDNSGYGY"
     terminator      8674..8721
                     /label=T7 term
                     /note="T7 terminator"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
     2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
     3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
     3061 cgttagcgag gtgccgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     3121 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagca
     3181 ataattttgt ttaactttaa gaaggagata tacatatggc taaagaaatt gcctttgacc
     3241 agaaatctag gagtgccctt caggctggta tcgataagct cgctgatgct gtcggtctca
     3301 ctcttggtcc cagggggagg aatgtagtgt tggatgaata tggcacccca aaggtggtta
     3361 atgacggagt tacaattgct cgtgctatag agctacctga tgctatggaa aatgctggcg
     3421 ctgcccttat cagggaggtt gcaagcaaaa ctaacgattc agccggtgat ggaaccacga
     3481 ctgcatctgt tcttgctcgc gaaatcatta aacttggtct attgagtgtt acatctggtg
     3541 caaatccagt gtctttgaag aagggcattg acaagactgt actgggtttg attgaagagc
     3601 tagaaaagaa ggctagacct gttaaaggtc gtgatgacat caaagctatt gcttcgatct
     3661 ctgctggaaa tgatgaaagc attggcacca tgattgcgga tgcaattgac aaagtcggtc
     3721 cagatggtgt gttgtccatc gagtcatcct cctcctttga gacaactgtt catgttgaag
     3781 aaggaatgga gattgataga ggatatattt ccccacaatt tgtcaccaac cctgagaaat
     3841 taattgttga gtttgagaat gctagagtct tagttacgga tcagaagatc tcggctatca
     3901 aggatattat ccccctattg gaaaagacaa ctcagttacg ctctcctctg ctcattattg
     3961 ctgaggatgt caccggggaa gctctggcta ctcttgtcgt gaacaagctg cggggtatac
     4021 tgaatgttgc cgccatcaaa gctcctggat ttggtgaaag gaggaaggct cttctgcaag
     4081 atattgccat tgtgacaggg gctgagtacc aagcaactga cctgggcctg ctcattgagg
     4141 gtaccccagt tgaagcactt ggaattgcca gaaaggtgac cattaccaag gattcaacaa
     4201 ccatcattgc tgatgcagca tcgaaggacg agatacaggc taggattgct cagctcaaaa
     4261 aggagttgtc cgagacagac tcggtttatg actccgagaa acttgctgag agaatcgcca
     4321 agttgtctgg gggtgttgcc gtcatcaagg tcggagctgc tacagaggct gagcttgagg
     4381 accgcaagct tcgcatcgag gatgcaaaga atgcaacttt tgctgcaatc gaagagggaa
     4441 tagtacctgg tggtggtgct gcctttgttc atctatcaac ttatgtccct gccattaagg
     4501 acaagctcga agatgcagat gaaaagttgg gagctgacat cattcaaagg gcattagtag
     4561 caccagcatc tttgatagcc caaaatgctg gaattgaagg ggaagtagta gtggagaagg
     4621 taagggaagc tgaatgggag atgggttata acgcgatgac agacaaatac gagaatcttg
     4681 ttgatgctgg agtgatcgat ccagccaagg tgacaagatg tgcattgcag aattcagcat
     4741 cagttgcagg aatggtactt acaacacaag ccattgtggt cgagaaggca aagcctaagg
     4801 cagctgcacc tgctgctcca caagggctct cagtgtgata cttggcgcgc ggataacaat
     4861 tgccataata cgactcacta tagggagacc acaacggttt ccctctagaa ataattttgt
     4921 ttaactttaa gaaggagata tacatatgaa tattcgtcca ttgcatgatc gcgtgatcgt
     4981 caagcgtaaa gaagttgaaa ctaaatctgc tggcggcatc gttctgaccg gctctgcagc
     5041 ggctaaatcc acccgcggcg aagtgctggc tgtcggcaat ggccgtatcc ttgaaaatgg
     5101 cgaagtgaag ccgctggatg tgaaagttgg cgacatcgtt attttcaacg atggctacgg
     5161 tgtgaaatct gagaagatcg acaatgaaga agtgttgatc atgtccgaaa gcgacattct
     5221 ggcaattgtt gaagcgtaat acttggcgcg cgaagttaat acgactcact ataggggaat
     5281 tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat
     5341 acatacccat gtcaccacaa acagagacta aagcaagtgt tggattcaaa gctggtgtta
     5401 aagagtacaa attgacttat tatactcctg agtaccaaac caaggatact gatatattgg
     5461 cagcattccg agtaactcct caacctggag ttccacctga agaagcaggg gccgcggtag
     5521 ctgccgaatc ttctactggt acatggacaa ctgtatggac cgatggactt accagccttg
     5581 atcgttacaa agggcgatgc taccgcatcg agcgtgttgt tggagaaaaa gatcaatata
     5641 ttgcttatgt agcttaccct ttagaccttt ttgaagaagg ttctgttacc aacatgttta
     5701 cttccattgt aggtaacgta tttgggttca aagccctgcg cgctctacgt ctggaagatc
     5761 tgcgaatccc tcctgcttat gttaaaactt tccaaggtcc gcctcatggg atccaagttg
     5821 aaagagataa attgaacaag tatggtcgtc ccctgttggg atgtactatt aaacctaaat
     5881 tggggttatc tgctaaaaac tacggtagag ctgtttatga atgtcttcgc ggtggacttg
     5941 attttaccaa agatgatgag aacgtgaact cacaaccatt tatgcgttgg agagatcgtt
     6001 tcttattttg tgccgaagca ctttataaag cacaggctga aacaggtgaa atcaaagggc
     6061 attacttgaa tgctactgca ggtacatgcg aagaaatgat caaaagagct gtatttgcta
     6121 gagaattggg cgttccgatc gtaatgcatg actacttaac ggggggattc accgcaaata
     6181 ctagcttggc tcattattgc cgagataatg gtctacttct tcacatccac cgtgcaatgc
     6241 atgcggttat tgatagacag aagaatcatg gtatccactt ccgggtatta gcaaaagcgt
     6301 tacgtatgtc tggtggagat catattcact ctggtaccgt agtaggtaaa cttgaaggtg
     6361 aaagagacat aactttgggc tttgttgatt tactgcgtga tgattttgtt gaacaagatc
     6421 gaagtcgcgg tatttatttc actcaagatt gggtctcttt accaggtgtt ctacccgtgg
     6481 cttcaggagg tattcacgtt tggcatatgc ctgctctgac cgagatcttt ggggatgatt
     6541 ccgtactaca gttcggtgga ggaactttag gacatccttg gggtaatgcg ccaggtgccg
     6601 tagctaatcg agtagctcta gaagcatgtg taaaagctcg taatgaagga cgtgatcttg
     6661 ctcaggaagg taatgaaatt attcgcgagg cttgcaaatg gagcccggaa ctagctgctg
     6721 cttgtgaagt atggaaagag atcgtattta attttgcagc agtggacgtt ttggataagt
     6781 aatacttggc gcgcccagtg cggccgcata atgcttaagt cgaacagaaa gtaatcgtat
     6841 tgtacacggc cgcataatcg aaattaatac gactcactat aggggaattg tgagcggata
     6901 acaattcccc atcttagtat attagttaag tataagaagg agatatacat atacatatgg
     6961 cagcaaagga gttgtatttc aacaaggatg gatcagctat caagaggctg caggttggtg
     7021 tgaacaaact cgctgatcta gttggagtta ctcttggccc aaaaggcagg aatgttgttc
     7081 ttgagagcaa gtatggtgct ccaaagattg tcaatgatgg agttactgtt gccagagagg
     7141 ttgagttaga ggatccagta gaaaacattg gtgctaaact agtgagacaa gctgctgcca
     7201 aaaccaatga tttggctggt gatgggacta caacatctgt cgtactagcc cagggcctgg
     7261 ttgctgaagg tgttaaggtg gttgcagctg gtgcaaaccc tgtcctgatc accagaggaa
     7321 ttgagaagac cacaaaagcc ttagtagctg aattgaagaa tatgtcaaaa gaggtagaag
     7381 acagtgaatt agcagatgtt gctgcagtaa gtgctgggaa caactctgaa gtagggagta
     7441 tgattgctga agccatgagc aaggttggaa ggaaaggtgt tgtgactctt gaagagggaa
     7501 aaagtgctga aaacagtctc cgtgtggttg aaggaatgca atttgaccgt ggttacatct
     7561 caccatactt cgttactgat agtgagaaaa tgtccgttga gtatgagaac tgtaagttat
     7621 tactggttga caagaagata acaaatgcaa gagatcttgt tggtgtcctg gaagatgcta
     7681 tcagaaatgg ttacccaatt ttaattattg ccgaagatat tgagcaagaa gcactagcca
     7741 cccttgttgt caataagctt agaggtgctt taaaggttgc tgcccttaaa gctcctggtt
     7801 ttggtgaacg aaaaagccag tatcttgatg acatagcaac ccttactgga ggcactgtga
     7861 ttagggagga acttggcctt accttagaca aggctgacaa ggaagttcta ggccatgctt
     7921 ctaaggtagt gctgactaag gatgccacca caattgttgg tgatggtagc actcaggaag
     7981 cagtcaataa gcgtgttgta cagattaaaa acctcataga ggcggcagat caagattatg
     8041 aaaaagaaaa gctaaatgaa agaattgcta aattatcagg aggtgtggct gttatacagg
     8101 ttggagcaca aactgaaact gaattgaagg aaaagaagct gagggtggaa gatgctctca
     8161 atgcaacaaa ggcagctgtt gaggaaggta ttgttgttgg tggagggtgc acactcctgc
     8221 gacttgcttc taaagttgat gccatcaagg agactcttga gaatgatgag gagaaggttg
     8281 gagctgatat tgttaaaaga gcgttgagct acccattgaa gttgattgct aaaaatgctg
     8341 gtgttaatgg aagtgttgtc agtgagaagg tgctctctag tgacaatcca aaatttggat
     8401 acaatgctgc aactggcaat tatgaagact tgatggctgc tggtattatt gacccaacaa
     8461 aggtggtgag atgttgtcta gagcatgcgt catcagtggc gaagacattc ttgatgtcag
     8521 attgtgtggt ggtagagatc aaggagcctg aaccagcagt tgctggcaac ccaatggaca
     8581 attcaggtta tggatactaa ttatgttata tctttgtaat ctgcaacgta ccgaaaggaa
     8641 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     8701 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc cgcaagaggc
     8761 ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga
     8821 tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact
     8881 gtgataaact accgcattaa agctagctta tcgatgataa gctgtcaaac atgagaa
//
