LOCUS       V009469                10160 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V009469
VERSION     V009469
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10160)
  AUTHORS   Kang MK, Eom JH, Kim Y, Um Y, Woo HM.
  TITLE     Biosynthesis of pinene from glucose using metabolically-engineered
            Corynebacterium glutamicum
  JOURNAL   Biotechnol. Lett. (2014) In press
   PUBMED   24930112
REFERENCE   2  (bases 1 to 10160)
  AUTHORS   Woo HM.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-MAY-2014) Clean Energy Research Center, Korea
            Institute of Science and Technology, Hwarangno 14-gil 5,
            Seongbuk-gu, Seoul 136-791, Korea
REFERENCE   3  (bases 1 to 10160)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10160)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Lett. (2014) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (13-MAY-2014) Clean Energy Research Center, Korea Institute of
            Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul
            136-791, Korea"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        266..294
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    302..318
                     /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)."
     CDS             370..2253
                     /gene="ag3"
                     /label="Pinene synthase, chloroplastic"
                     /note="Pinene synthase, chloroplastic from Abies grandis.
                     Accession#: O24475"
     RBS             2263..2274
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     CDS             2281..3432
                     /codon_start=1
                     /product="geranyl diphosphate synthase"
                     /label="geranyl diphosphate synthase"
                     /note="prenyltransferase AgGPPS1; codon-optimized for C.
                     glutamicum"
                     /protein_id="AIF29195.1"
                     /translation="MAYSCMAASCHGLHFMNIASQECNLKRGIIPSKRLHGISSSLWAS
                     NGFQGHLERDLSAYRHLVSSSRCLNTIAMLSNLSEQAKEKATEFDFKEYLHSKAISVNE
                     ALERAVPLRYPEKIHEAMRYSLLAGGKRIRPILTIAACELVGGSEELAMPTACAMEMIH
                     TMSLIHDDLPSMDNDDLRRGKLTNHKVFGEGTAVLAGDALLSFAFEHIAVSTRKTVASH
                     RVLRVVSELGKAIGSQGVAGGQVADITSEGDPSVGLETLEWIHIHKTAVLLECAVVSGA
                     IIGGASENEIERTGRYARCVGLLFQVVDDILDVTRSSEELGKTAGKDLVSDKATYPKLM
                     GLEKAKEFADELLDRAKEELSCFNPAKAAPLLGLADYIALRQN"
     terminator      3659..3745
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3837..3864
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3883..3974
                     /label="AmpR promoter"
     CDS             complement(7922..8734)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(9329..9874)
                     /direction=LEFT
                     /label="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin."
ORIGIN
        1 cggtacgcct gcggccaaat ttgattcctt tcagctttgc ttcctgtcgg ccctcattcg
       61 tgcgctctag gatcagtgat ccgggcttat cgactgcacg gtgcaccaat gcttctggcg
      121 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
      181 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
      241 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagaattc aaaagatcta ttaaagagga
      361 gaatactaga tggcactggt ttccaccgca ccactggcat ccaagtcctg cctgcacaag
      421 tccctgatct cctccaccca cgaactgaag gcactgtccc gcaccatccc agcactgggc
      481 atgtcccgcc gcggcaagtc catcacccca tccatctcca tgtcctccac caccgtcgtc
      541 accgacgacg gcgtccgccg ccgcatgggc gacttccact ccaacctgtg ggacgacgac
      601 gtcatccagt ccctgccaac cgcatacgaa gaaaagtcct acctggaacg cgcagaaaag
      661 ctgatcggcg aagtcaagaa catgttcaac tccatgtccc tggaagacgg cgaactgatg
      721 tccccactga acgacctgat ccagcgcctg tggatcgtcg actccctgga acgcctgggc
      781 atccaccgcc acttcaagga cgaaatcaag tccgcactgg actacgtcta ctcctactgg
      841 ggcgaaaacg gcatcggctg cggccgcgaa tccgtcgtca ccgacctgaa ctccaccgca
      901 ctgggcctgc gcaccctgcg cctgcacggc tacccagtct cctccgacgt cttcaaggca
      961 ttcaagggcc agaacggcca gttctcctgc tccgaaaaca tccagaccga cgaagaaatc
     1021 cgcggcgtcc tgaacctgtt ccgcgcatcc ctgatcgcat tcccaggcga aaagatcatg
     1081 gacgaagcag aaatcttctc caccaagtac ctgaaggaag cactgcaaaa gatcccagtc
     1141 tcctccctgt cccgcgaaat cggcgacgtc ctggaatacg gctggcacac ctacctgcca
     1201 cgcctggaag cacgcaacta catccaggtc ttcggccagg acaccgaaaa caccaagtcc
     1261 tacgtcaagt ccaagaagct gctggaactg gcaaagctgg agttcaacat cttccagtcc
     1321 ctccagaagc gcgaactgga atccctggtc cgctggtgga aggaatccgg cttcccagaa
     1381 atgaccttct gccgccaccg ccacgtcgaa tactacaccc tggcatcctg catcgcattc
     1441 gaaccacagc actccggctt ccgcctgggc ttcgcaaaga cctgccacct gatcaccgtc
     1501 ctggacgaca tgtacgacac cttcggcacc gtcgacgaac tggaactgtt caccgcaacc
     1561 atgaagcgct gggacccatc ctccatcgac tgcctgccag aatacatgaa gggcgtctac
     1621 atcgcagtct acgacaccgt caacgaaatg gcacgcgaag cagaagaagc acagggccgc
     1681 gacaccctga cctacgcacg cgaagcatgg gaagcataca tcgactccta catgcaggaa
     1741 gcacgctgga tcgcaaccgg ctacctgcca tccttcgacg aatactacga aaacggcaag
     1801 gtttcctgcg gccaccgcat ctccgcactc cagccaatcc tgaccatgga catcccattc
     1861 ccagaccaca tcctgaagga agtcgacttc ccatccaagc tgaacgacct ggcatgcgca
     1921 atcctgcgcc tgcgcggcga cacccgctgc tacaaggcag accgcgcacg cggcgaagaa
     1981 gcatcctcca tctcctgcta catgaaggac aacccaggcg tctccgaaga agacgcactg
     2041 gaccacatca acgcaatgat ctccgacgtc atcaagggcc tgaactggga actgctgaag
     2101 ccagacatca acgtcccaat ctccgcaaag aagcacgcat tcgacatcgc acgcgcattc
     2161 cactacggct acaagtaccg cgacggctac tccgtcgcaa acgtcgaaac caagtccctg
     2221 gtcacccgca ccctgctgga atccgtccca ctgtaaggat ctaaagagga gaaatactag
     2281 atggcatact cctgcatggc agcatcctgc cacggcctgc acttcatgaa catcgcatcc
     2341 caggaatgca acctgaagcg cggcatcatc ccatccaagc gcctgcacgg catctcctcc
     2401 tccctgtggg catccaacgg cttccagggc cacctggaac gcgacctgtc cgcataccgc
     2461 cacctggttt cctcctcccg ctgcctgaac accatcgcaa tgctgtccaa cctgtccgaa
     2521 caggcaaagg aaaaggcaac cgagttcgac ttcaaggaat acctgcactc caaggcaatc
     2581 tccgtcaacg aagcactgga acgcgcagtc ccactgcgct acccagaaaa gatccacgaa
     2641 gcaatgcgct actccctgct ggcaggcggc aagcgcatcc gcccaatcct gaccatcgca
     2701 gcatgcgaac tggtcggcgg ctccgaagaa ctggcaatgc caaccgcatg cgcaatggaa
     2761 atgatccaca ccatgtccct gatccacgac gacctgccat ccatggacaa cgacgacctg
     2821 cgccgcggca agctgaccaa ccacaaggtc ttcggcgaag gcaccgcagt cctggcaggc
     2881 gacgcactgc tgtccttcgc attcgaacac atcgcagtct ccacccgcaa gaccgtcgca
     2941 tcccaccgcg tcctgcgcgt cgtctccgaa ctgggcaagg caatcggctc ccagggcgtc
     3001 gcaggcggcc aggtcgcaga catcacctcc gaaggcgacc catccgtcgg cctggaaacc
     3061 ctggaatgga ttcacatcca caagaccgca gtcctgctgg aatgcgcagt cgtctccggc
     3121 gcaatcatcg gcggcgcatc cgaaaacgaa atcgaacgca ccggccgcta cgcacgctgc
     3181 gtcggcctgc tgttccaggt cgtcgacgac atcctggacg tcacccgctc ctccgaagaa
     3241 ctgggcaaga ccgcaggcaa ggacctggtt tccgacaagg caacctaccc aaagctgatg
     3301 ggcctggaaa aggcaaagga gttcgcagac gaactgctgg accgcgcaaa ggaagaactg
     3361 tcctgcttca acccagcaaa ggcagcacca ctgctgggcc tggcagacta catcgcactg
     3421 cgccagaact aaggatccgt cgacctgcag ccaagcttct gttttggcgg atgagagaag
     3481 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg
     3541 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc
     3601 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca
     3661 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     3721 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
     3781 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
     3841 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa
     3901 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3961 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4021 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4081 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4141 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4201 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4261 ctcagaatga cttggttgag tactgcggcg tcgctgatcg ccctggcgac gttgtgcggg
     4321 tggcttgtcc ctgagggcgc tgcgacagat agctaaaaat ctgggtcagg atcgccgtag
     4381 agcgcgcgtc gtcgattgga ggcttcccct ttggttgacg gtcttcaatc gctctacggc
     4441 gatcctgacg cttttttgtt gcgtaccgtc gatcgtttta tttctgtcga tcccgaaaaa
     4501 gtttttgcct tttgtaaaaa acttctcggt cgccccgcaa attttcgatt ccagattttt
     4561 taaaaaccaa gccagaaata cgacacaccg tttgcagata atctgtcttt cggaaaaatc
     4621 aagtgcgata caaaattttt agcacccctg agctgcgcaa agtcccgctt cgtgaaaatt
     4681 ttcgtgccgc gtgattttcc gccaaaaact ttaacgaacg ttcgttataa tggtgtcatg
     4741 accttcacga cgaagtacta aaattggccc gaatcatcag ctatggatct ctctgatgtc
     4801 gcgctggagt ccgacgcgct cgatgctgcc gtcgatttaa aaacggtgat cggatttttc
     4861 cgagctctcg atacgacgga cgcgccagca tcacgagact gggccagtgc cgcgagcgac
     4921 ctagaaactc tcgtggcgga tcttgaggag ctggctgacg agctgcgtgc tcggccagcg
     4981 ccaggaggac gcacagtagt ggaggatgca atcagttgcg cctactgcgg tggcctgatt
     5041 cctccccggc ctgacccgcg aggacggcgc gcaaaatatt gctcagatgc gtgtcgtgcc
     5101 gcagccagcc gcgagcgcgc caacaaacgc cacgccgagg agctggaggc ggctaggtcg
     5161 caaatggcgc tggaagtgcg tcccccgagc gaaattttgg ccatggtcgt cacagagctg
     5221 gaagcggcag cgagaattat ccgcgatcgt ggcgcggtgc ccgcaggcat gacaaacatc
     5281 gtaaatgccg cgtttcgtgt ggccgtggcc gcccaggacg tgtcagcgcc gccaccacct
     5341 gcaccgaatc ggcagcagcg tcgcgcgtcg aaaaagcgca caggcggcaa gaagcgataa
     5401 gctgcacgaa tacctgaaaa atgttgaacg ccccgtgagc ggtaactcac agggcgtcgg
     5461 ctaaccccca gtccaaacct gggagaaagc gctcaaaaat gactctagcg gattcacgag
     5521 acattgacac accggcctgg aaattttccg ctgatctgtt cgacacccat cccgagctcg
     5581 cgctgcgatc acgtggctgg acgagcgaag accgccgcaa attcctcgct cacctgggca
     5641 gagaaaattt ccagggcagc aagacccgcg acttcgccag cgcttggatc aaagacccgg
     5701 acacgggaga aacacagccg aagttatacc gagttggttc aaaatcgctt gcccggtgcc
     5761 agtatgttgc tctgacgcac gcgcagcacg cagccgtgct tgtcctggac attgatgtgc
     5821 cgagccacca ggccggcggg aaaatcgagc acgtaaaccc cgaggtctac gcgattttgg
     5881 agcgctgggc acgcctggaa aaagcgccag cttggatcgg cgtgaatcca ctgagcggga
     5941 aatgccagct catctggctc attgatccgg tgtatgccgc agcaggcatg agcagcccga
     6001 atatgcgcct gctggctgca acgaccgagg aaatgacccg cgttttcggc gctgaccagg
     6061 ctttttcaca taggctgagc cgtggccact gcactctccg acgatcccag ccgtaccgct
     6121 ggcatgccca gcacaatcgc gtggatcgcc tagctgatct tatggaggtt gctcgcatga
     6181 tctcaggcac agaaaaacct aaaaaacgct atgagcagga gttttctagc ggacgggcac
     6241 gtatcgaagc ggcaagaaaa gccactgcgg aagcaaaagc acttgccacg cttgaagcaa
     6301 gcctgccgag cgccgctgaa gcgtctggag agctgatcga cggcgtccgt gtcctctgga
     6361 ctgctccagg gcgtgccgcc cgtgatgaga cggcttttcg ccacgctttg actgtgggat
     6421 accagttaaa agcggctggt gagcgcctaa aagacaccaa gggtcatcga gcctacgagc
     6481 gtgcctacac cgtcgctcag gcggtcggag gaggccgtga gcctgatctg ccgccggact
     6541 gtgaccgcca gacggattgg ccgcgacgtg tgcgcggcta cgtcgctaaa ggccagccag
     6601 tcgtccctgc tcgtcagaca gagacgcaga gccagccgag gcgaaaagct ctggccacta
     6661 tgggaagacg tggcggtaaa aaggccgcag aacgctggaa agacccaaac agtgagtacg
     6721 cccgagcaca gcgagaaaaa ctagctaagt ccagtcaacg acaagctagg aaagctaaag
     6781 gaaatcgctt gaccattgca ggttggttta tgactgttga gggagagact ggctcgtggc
     6841 cgacaatcaa tgaagctatg tctgaattta gcgtgtcacg tcagaccgtg aatagagcac
     6901 ttaaggtctg cgggcattga acttccacga ggacgccgaa agcttcccag taaatgtgcc
     6961 atctcgtagg cagaaaacgg ttcccccgta gggtctctct cttggcctcc tttctaggtc
     7021 gggctgattg ctcttgaagc tctctagggg ggctcacacc ataggcagat aacgttcccc
     7081 accggctcgc ctcgtaagcg cacaaggact gctcccaaag atcttcaaag ccactgccgc
     7141 gactgccttc gcgaagcctt gccccgcgga aatttcctcc accgagttcg tgcacacccc
     7201 tatgccaagc ttctttcacc ctaaattcga gagattggat tcttaccgtg gaaattcttc
     7261 gcaaaaatcg tcccctgatc gcccttgcga cgttggcgtc ggtgccgctg gttgcgcttg
     7321 gcttgaccga cttgatcctc cggcgttcag cctgtgccac agccgacagg atggtgacca
     7381 ccatttgccc catatcaccg tcggtactga tcccgtcgtc aataaaccga accgctacac
     7441 cctgagcatc aaactctttt atcagttgga tcatgtcggc gtgtcgcggc caagacggtc
     7501 gagcttcttc accagaatga catcaccttc ctccaccttc atcctcagca aatccagccc
     7561 ttcccgatct gttgaactgc cggatgcctt gtcggtaaag atgcggttag cttttacccc
     7621 tgcatctttg agcgctgagg tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc
     7681 tgaatcgccc catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg
     7741 taggtggacc agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg
     7801 ggaagatgcg tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccgc
     7861 cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt
     7921 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     7981 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     8041 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     8101 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     8161 aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc
     8221 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     8281 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     8341 gcaaccggcg caggaacact gccagcgcat cagcaatatt ttcacctgaa tcaggatatt
     8401 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     8461 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     8521 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     8581 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     8641 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     8701 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     8761 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     8821 gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
     8881 agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca
     8941 ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg
     9001 atgatggggc gattcaggcc tggtatgagt cagcaacacc ttcttcacga ggcagacctc
     9061 agcgctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt
     9121 gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg
     9181 atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg
     9241 gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg
     9301 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     9361 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     9421 cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     9481 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag
     9541 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc
     9601 ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca
     9661 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta
     9721 aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga
     9781 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga
     9841 gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta aggggtctga
     9901 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     9961 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    10021 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    10081 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    10141 gggcttacca tctggcccca
//
