LOCUS       V002911                 8289 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002911
VERSION     V002911
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8289)
  AUTHORS   Lu Y, Rijzaani H, Karcher D, Ruf S, Bock R.
  TITLE     Efficient engineering of the vitamin E metabolic pathway in
            transgenic tobacco and tomato plastids with synthetic multigene
            operons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8289)
  AUTHORS   Lu Y, Rijzaani H, Karcher D, Ruf S, Bock R.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2012) Organelle Biology, Biotechnology and
            Molecular Ecophysiology, Max Planck Institute of Molecular Plant
            Physiology, Am Muehlenberg 1, Potsdam-Golm, Brandenburg 14476,
            Germany
REFERENCE   3  (bases 1 to 8289)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8289)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (25-JUN-2012) Organelle Biology, Biotechnology and Molecular
            Ecophysiology, Max Planck Institute of Molecular Plant Physiology,
            Am Muehlenberg 1, Potsdam-Golm, Brandenburg 14476, Germany"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     tRNA            complement(31..104)
                     /product="tRNA-Met"
     CDS             complement(257..556)
                     /gene="rps14"
                     /label="Small ribosomal subunit protein uS14c"
                     /note="Small ribosomal subunit protein uS14c from Nicotiana
                     sylvestris. Accession#: Q3C1I0"
     gene            complement(679..1878)
                     /gene="psaB"
                     /label="psaB"
                     /note="derived from Nicotiana tabacum"
     promoter        complement(1891..1909)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1919..1935)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      2077..2532
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2558..2662
                     /label="AmpR promoter"
     CDS             2663..3520
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      3694..4282
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    4570..4591
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4606..4636
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4644..4660
                     /label="lac operator"
                     /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)."
     primer_bind     4668..4684
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        4705..4723
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     gene            4737..4867
                     /gene="psbZ"
                     /label="psbZ"
                     /note="derived from Nicotiana tabacum"
     tRNA            5143..5213
                     /product="tRNA-Gly"
     protein_bind    5406..5439
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      5480..5614
                     /note="prrN promoter; derived from Nicotiana tabacum"
                     /regulatory_class="promoter"
     CDS             5615..6403
                     /label="SmR"
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      6417..6811
                     /note="psbA terminator; derived from Nicotiana tabacum"
                     /regulatory_class="terminator"
     misc_recomb     6815..6848
                     /label="LoxP"
                     /note="LoxP"
     protein_bind    complement(6815..6848)
                     /label="loxP"
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
     primer_bind     6878..6894
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      complement(6897..7117)
                     /note="rbcL terminator; derived from Nicotiana tabacum"
                     /regulatory_class="terminator"
     CDS             complement(7118..8071)
                     /codon_start=1
                     /gene="TMT"
                     /product="gamma-tocopherol methyltransferase"
                     /label="TMT"
                     /note="derived from Synechocystis sp. PCC 6803"
                     /protein_id="AFQ99316.1"
                     /translation="MVYHVRPKHALFLAFYCYFSLLTMASATIASADLYEKIKNFYDDS
                     SGLWEDVWGEHMHHGYYGPHGTYRIDRRQAQIDLIKELLAWAVPQNSAKPRKILDLGCG
                     IGGSSLYLAQQHQAEVMGASLSPVQVERAGERARALGLGSTCQFQVANALDLPFASDSF
                     DWVWSLESGEHMPNKAQFLQEAWRVLKPGGRLILATWCHRPIDPGNGPLTADERRHLQA
                     IYDVYCLPYVVSLPDYEAIARECGFGEIKTADWSVAVAPFWDRVIESAFDPRVLWALGQ
                     AGPKIINAALCLRLMKWGYERGLVRFGLLTGIKPLV"
     gene            complement(7118..8071)
                     /gene="TMT"
                     /label="TMT"
                     /note="SyTMT"
     RBS             complement(8079..8101)
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(8236..8254)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aaccctaaaa tagtttggca aaacaagatg ttgcggagac aggatttgaa cccgtgacct
       61 caaggttatg agccttgcga gctaccaaac tgctctaccc cgcgccgaag ataagaactg
      121 aaaactaata gataaacaag gattaaatgc gcccctccac cctatctgta caaatagaat
      181 agcccattta tacagaatgg taaaggggct tctatgatca tcgaccatag aaatagaaat
      241 gaagcgttaa tccttaccaa cttgatcttg ttgctcctgg caacaaacat gcatgaacca
      301 tttcacgaag tatgtgtccg gatagtccaa agtctcgata gttagctctc ggccttccgg
      361 tcaaaaaaca acgtcgatga aggcgtgtag gtgcactatt ccgtggtggg gattgtaact
      421 ttccataaat ttcccatttg tcactcaacg acggaacctt gcttatttct ttctttgagg
      481 atcgacgaat cgaatgatat ttctgttcca atttttgcct cttcttctcc ctctgaatca
      541 aacttttcct tgccataatg gttgaattcc tattagtatc catgatacaa gtcgaatcct
      601 agatgtagaa atagaagaag gtggaccccc tctccgtcga aagaaatgag attatcgcag
      661 atacacacat taaaaatatt aaccaaattt gcccgacgta gaggcaatca agaaagccgc
      721 ataagtgaat atataaccta cagaaaagtg agctaatcca accaatcttg cttgtacaat
      781 ggaaagggcc actggtttat ctctccagcg aatcaaattg gccaaaggtg tgcgttcatg
      841 agcccatgct aaagtttcaa tcaattcctg ccaatatcca cgccaagaaa ttaagaacat
      901 aaatccagta gcccaaacaa gatgtccaaa taagaacatc catgcccaaa ccgataaact
      961 attcatacca aaaggattat atccgttgat aagttgtgaa gagtttaacc ataaataatc
     1021 ccttaaccag cccatcaaat aagtggaaga ttcattaaac tgtgaaacgt taccctgcca
     1081 taatgtgatg tgcttccaat gccaataaaa agtaacccat ccaatagtat ttaacatcca
     1141 aaaaactgcc aaataaaacg cgtcccatgc cgaaatatca caagtaccgc ctcgtcctgg
     1201 gccatcgcac ggaaaactat aaccgaaatc ctttttatct ggcattaact tggaaccacg
     1261 tgcatctaaa gcacctttta ctaagatcaa tgtagttgta tgtaaaccaa gagcaatagc
     1321 atgatgaacc aaaaagtctc caggacctat tgttaaaaat aatgaattac tattttcatt
     1381 aacagcattt aaccaacccg gcaaccagat gcttcgaccc gcattgaatg ctggaccact
     1441 cgttgaagat aaaagtacat cgaacccata tgaagtttta ccatgagcgg attgtatcca
     1501 ttgagcaaat ataggttcaa tcaagatttg cttctccgga gtgccaaagg caagcatgac
     1561 atcattatga acataaagtc ccagggtatg gaatcccaga aagaggctgg cccaacttaa
     1621 atgagatatg atagcttctt tatgctctaa cattcttgcc aatacattat cttcattttg
     1681 ctccggattg taatctctaa tgaaaaatat agctccatga gcaaaagctc ctgtcatgat
     1741 gaatcctgcg atatattggt ggtgggtata taatgcagct tgagtagtaa agtcttgtgc
     1801 tatgaatgca taagcaggta aagagtacat gtgttgagct accaaagaag taataacccc
     1861 taaagaagct agagcaaggc tccaattcgc cctatagtga gtcgtattac gcgcgctcac
     1921 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     1981 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     2041 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     2101 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2161 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2221 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2281 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2341 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2401 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     2461 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2521 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2581 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2641 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2701 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2761 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2821 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2881 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2941 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3001 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3061 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3121 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3181 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3241 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3301 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3361 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3421 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3481 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3541 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3601 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3661 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3721 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3781 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3841 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3901 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3961 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4021 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4081 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4141 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4201 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4261 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4321 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4381 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4441 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4501 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     4561 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     4621 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     4681 tgaccatgat tacgccaagc tcgaaattaa ccctcactaa agggaacaaa agctggtgat
     4741 tagcgtaccc gttgtatttg cttctcctga tggctggtca agtaacaaaa atgttgtatt
     4801 ttctggtaca tccttatgga ttggattagt ctttctggtg ggtatcctta attctctcat
     4861 ctcttgaacc tattcgtcgc agacccaaaa ccaaaatgac ccccctaatt tttctcggtt
     4921 gtgagacaca ttaaattgga atctaagtcc ccaaagaaaa cgcaaatcaa ataaagaaaa
     4981 caaaaaaatt agaggggggt caaacttctt gaataaaaag aatacaatta aaaaaataat
     5041 tggaatcgtt ccgaagagaa tatgtgtccc ggcactgcac aaaaaagatc cggttatata
     5101 tcatatatgt gggtacatat tgtgtatcaa gaacaaaaaa atgcggatat ggtcgaatgg
     5161 taaaatttct ctttgccaag gagaagatgc gggttcgatt cccgctatcc gcccaagatc
     5221 caagataaag taattttatt actatttatt tattatttaa tttcataaat agcattaaat
     5281 atatccttaa attaaggatt tggtatagtt ggccgtgata gtgtagtgat tctatccctc
     5341 ccctacgttt tctttttcct tccaccccca aaaagcgaaa ggcgggaatt aattactagc
     5401 taggtataac ttcgtataat gtatgctata cgaagttatt aattaatcga cagatcccga
     5461 attgggatcg gtacccaaag ctcccccgcc gtcgttcaat gagaatggat aagaggctcg
     5521 tgggattgac gtgagggggc agggatggct atatttctgg gagcgaactc cgggcgaata
     5581 cgaagcgctt ggatacagtt gtagggaggg atttatggca gaagcggtga tcgccgaagt
     5641 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     5701 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     5761 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     5821 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     5881 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     5941 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     6001 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     6061 ggcggaggaa ctctttgatc cggttcttga acaggatcta tttgaggcgc taaatgaaac
     6121 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     6181 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     6241 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     6301 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     6361 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc tagagcgatc
     6421 ctggcctagt ctataggagg ttttgaaaag aaaggagcaa taatcatttt cttgttctat
     6481 caagagggtg ctattgctcc tttctttttt tctttttatt tatttactag tattttactt
     6541 acatagactt ttttgtttac attatagaaa aagaaggaga ggttattttc ttgcatttat
     6601 tcatgattga gtattctatt ttgattttgt atttgtttaa aattgtagaa atagaacttg
     6661 tttctcttct tgctaatgtt actatatctt tttgattttt tttttccaaa aaaaaaatca
     6721 aattttgact tcttcttatc tcttatcttt gaatatctct tatctttgaa ataataatat
     6781 cattgaaata agaaagaaga gctatattcg acctataact tcgtataatg tatgctatac
     6841 gaagttattg aattctagtt agtaccgggc cccccctcga ggtcgacggt atcgataagc
     6901 ttgctagatt ttgtatttca aatcttgtat atctaggtaa gtatatactt agtcaaaata
     6961 tatgcaatag aatctttgtt gtattcggct caatcctttt agtaaaagat tgggccgagt
     7021 ttaattgcaa ttcaattaag agaacgaagg ataattactt gagttctttc tccttatcct
     7081 tctttatttc ctgctaattt atctgctaat gtctactcta gactaaaggc tttatccccg
     7141 ttaataagcc aaaacgcact aatccccgtt catagcccca tttcattaat cgtaaacaca
     7201 gggcggcatt gataattttt ggccccgctt gccccaaggc ccacaacacc cggggatcga
     7261 acgcagactc aatcacccgg tcccaaaaag gtgccaccgc cactgaccaa tcggcagtct
     7321 taatttcccc aaacccacat tccctggcga tcgcctcgta gtccggcagg gaaaccacat
     7381 agggcaaaca gtaaacgtca tagatggctt ggagatggcg acgttcatcg gcagtcaggg
     7441 ggccattgcc gggatcaatg ggacgatgac accaggtcgc taaaatcaga cggccacctg
     7501 gtttaagtac ccgccaagct tcttgtaaaa actgagcttt gttgggcatg tgctccccac
     7561 tttccaacga ccaaacccag tcaaaggaat cggaagcaaa gggcaaatcc aaggcattgg
     7621 ccacctgaaa ctggcaggtt gagcccaacc ccagggccct ggccctttcc cccgcccgtt
     7681 ccacctgcac tggggaaaga ctagccccca tcacttctgc ttggtgttgc tgggccaagt
     7741 acaaactact gccgccaatg ccacagccta aatcgagaat ttttcgtggt ttggcgctat
     7801 tttggggcac tgcccaggcc aatagttctt tgatcagatc aatttgagcc tggcggcgat
     7861 cgatccgata ggtgccgtgg ggaccgtagt agccgtggtg catatgctca ccccaaacgt
     7921 cttcccagag accgctggag tcgtcgtaga aatttttaat tttttcgtag aggtctgcac
     7981 tggcaatggt ggcgctggcc atggtaagca aagagaaata acaatagaat gctaagaaca
     8041 gggcgtgctt aggcctaaca tggtaaacca tatgtatatc tccttcttaa agttaaacaa
     8101 aattatttct agtgggaaac cgttgtggtc tccctcccag aaatatagcc atccctgccc
     8161 cctcacgtca atcccacgag cctcttatcc attctcattg aacgacggcg ggggagcgag
     8221 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     8281 gctgtttcc
//
