LOCUS       V009468                10154 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V009468
VERSION     V009468
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10154)
  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 10154)
  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 10154)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10154)
  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..10154
                     /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..3426
                     /codon_start=1
                     /product="geranyl diphosphate synthase"
                     /label="geranyl diphosphate synthase"
                     /note="AgGPPS2; codon-optimized for C. glutamicum"
                     /protein_id="AIF29189.1"
                     /translation="MAYSAMATMGYNGMAASCHTLHPTSPLKPFHGASTSLEAFNGEHM
                     GLLRGYSKRKLSSYKNPASRSSNATVAQLLNPPQKGKKAVEFDFNKYMDSKAMTVNEAL
                     NKAIPLRYPQKIYESMRYSLLAGGKRVRPVLCIAACELVGGTEELAIPTACAIEMIHTM
                     SLMHDDLPCIDNDDLRRGKPTNHKIFGEDTAVTAGNALHSYAFEHIAVSTSKTVGADRI
                     LRMVSELGRATGSEGVMGGQMVDIASEGDPSIDLQTLEWIHIHKTAMLLECSVVCGAII
                     GGASEIVIERARRYARCVGLLFQVVDDILDVTKSSDELGKTAGKDLISDKATYPKLMGL
                     EKAKEFSDELLNRAKGELSCFDPVKAAPLLGLADYVAFRQN"
     terminator      3653..3739
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3831..3858
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3877..3968
                     /label="AmpR promoter"
     CDS             complement(7916..8728)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(9323..9868)
                     /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 ccgcaatggc aaccatgggc tacaacggca tggcagcatc ctgccacacc
     2341 ctgcacccaa cctccccact gaagccattc cacggcgcat ccacctccct ggaagcattc
     2401 aacggcgaac acatgggcct gctgcgcggc tactccaagc gcaagctgtc ctcctacaag
     2461 aacccagcat cccgctcctc caacgcaacc gtggcacagc tgctgaaccc accacagaag
     2521 ggcaagaagg cagtggagtt cgatttcaac aagtacatgg attccaaggc aatgaccgtg
     2581 aacgaagcac tgaacaaggc aatcccactg cgctacccac agaaaatcta cgaatccatg
     2641 cgctactccc tgctggcagg cggcaagcgc gtgcgcccag tgctgtgcat cgcagcatgc
     2701 gaactggtgg gcggcaccga agaactggca atcccaaccg catgcgcaat cgaaatgatc
     2761 cacaccatgt ccctgatgca cgatgatctg ccatgcatcg ataacgatga tctgcgccgc
     2821 ggcaagccaa ccaaccacaa gattttcggc gaagataccg cagtgaccgc aggcaacgca
     2881 ctgcactcct acgcattcga acacatcgca gtgtccacct ccaagaccgt gggcgcagat
     2941 cgcatcctgc gcatggtgtc cgaactgggc cgcgcaaccg gctccgaagg cgtgatgggc
     3001 ggccagatgg tggatatcgc atccgaaggc gatccatcca tcgatctgca gaccctggaa
     3061 tggattcaca tccacaagac cgcaatgctg ctggaatgct ccgtggtgtg cggcgcaatc
     3121 atcggcggcg catccgaaat cgtgatcgaa cgcgcacgcc gctacgcacg ctgcgtgggc
     3181 ctgctgttcc aggtggtgga tgatatcctg gatgtgacca agtcctccga tgaactgggc
     3241 aagaccgcag gcaaggatct gatctccgat aaggcaacct acccaaagct gatgggcctg
     3301 gaaaaggcaa aggaattttc cgatgaactg ctgaaccgcg caaagggcga actgtcctgc
     3361 ttcgatccag tgaaggcagc accactgctg ggcctggcag attacgtggc attccgccag
     3421 aactaaggat ccgtcgacct gcagccaagc ttctgttttg gcggatgaga gaagattttc
     3481 agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc
     3541 ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc
     3601 gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa
     3661 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     3721 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     3781 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat
     3841 cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt
     3901 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3961 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4021 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4081 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4141 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4201 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4261 atgacttggt tgagtactgc ggcgtcgctg atcgccctgg cgacgttgtg cgggtggctt
     4321 gtccctgagg gcgctgcgac agatagctaa aaatctgggt caggatcgcc gtagagcgcg
     4381 cgtcgtcgat tggaggcttc ccctttggtt gacggtcttc aatcgctcta cggcgatcct
     4441 gacgcttttt tgttgcgtac cgtcgatcgt tttatttctg tcgatcccga aaaagttttt
     4501 gccttttgta aaaaacttct cggtcgcccc gcaaattttc gattccagat tttttaaaaa
     4561 ccaagccaga aatacgacac accgtttgca gataatctgt ctttcggaaa aatcaagtgc
     4621 gatacaaaat ttttagcacc cctgagctgc gcaaagtccc gcttcgtgaa aattttcgtg
     4681 ccgcgtgatt ttccgccaaa aactttaacg aacgttcgtt ataatggtgt catgaccttc
     4741 acgacgaagt actaaaattg gcccgaatca tcagctatgg atctctctga tgtcgcgctg
     4801 gagtccgacg cgctcgatgc tgccgtcgat ttaaaaacgg tgatcggatt tttccgagct
     4861 ctcgatacga cggacgcgcc agcatcacga gactgggcca gtgccgcgag cgacctagaa
     4921 actctcgtgg cggatcttga ggagctggct gacgagctgc gtgctcggcc agcgccagga
     4981 ggacgcacag tagtggagga tgcaatcagt tgcgcctact gcggtggcct gattcctccc
     5041 cggcctgacc cgcgaggacg gcgcgcaaaa tattgctcag atgcgtgtcg tgccgcagcc
     5101 agccgcgagc gcgccaacaa acgccacgcc gaggagctgg aggcggctag gtcgcaaatg
     5161 gcgctggaag tgcgtccccc gagcgaaatt ttggccatgg tcgtcacaga gctggaagcg
     5221 gcagcgagaa ttatccgcga tcgtggcgcg gtgcccgcag gcatgacaaa catcgtaaat
     5281 gccgcgtttc gtgtggccgt ggccgcccag gacgtgtcag cgccgccacc acctgcaccg
     5341 aatcggcagc agcgtcgcgc gtcgaaaaag cgcacaggcg gcaagaagcg ataagctgca
     5401 cgaatacctg aaaaatgttg aacgccccgt gagcggtaac tcacagggcg tcggctaacc
     5461 cccagtccaa acctgggaga aagcgctcaa aaatgactct agcggattca cgagacattg
     5521 acacaccggc ctggaaattt tccgctgatc tgttcgacac ccatcccgag ctcgcgctgc
     5581 gatcacgtgg ctggacgagc gaagaccgcc gcaaattcct cgctcacctg ggcagagaaa
     5641 atttccaggg cagcaagacc cgcgacttcg ccagcgcttg gatcaaagac ccggacacgg
     5701 gagaaacaca gccgaagtta taccgagttg gttcaaaatc gcttgcccgg tgccagtatg
     5761 ttgctctgac gcacgcgcag cacgcagccg tgcttgtcct ggacattgat gtgccgagcc
     5821 accaggccgg cgggaaaatc gagcacgtaa accccgaggt ctacgcgatt ttggagcgct
     5881 gggcacgcct ggaaaaagcg ccagcttgga tcggcgtgaa tccactgagc gggaaatgcc
     5941 agctcatctg gctcattgat ccggtgtatg ccgcagcagg catgagcagc ccgaatatgc
     6001 gcctgctggc tgcaacgacc gaggaaatga cccgcgtttt cggcgctgac caggcttttt
     6061 cacataggct gagccgtggc cactgcactc tccgacgatc ccagccgtac cgctggcatg
     6121 cccagcacaa tcgcgtggat cgcctagctg atcttatgga ggttgctcgc atgatctcag
     6181 gcacagaaaa acctaaaaaa cgctatgagc aggagttttc tagcggacgg gcacgtatcg
     6241 aagcggcaag aaaagccact gcggaagcaa aagcacttgc cacgcttgaa gcaagcctgc
     6301 cgagcgccgc tgaagcgtct ggagagctga tcgacggcgt ccgtgtcctc tggactgctc
     6361 cagggcgtgc cgcccgtgat gagacggctt ttcgccacgc tttgactgtg ggataccagt
     6421 taaaagcggc tggtgagcgc ctaaaagaca ccaagggtca tcgagcctac gagcgtgcct
     6481 acaccgtcgc tcaggcggtc ggaggaggcc gtgagcctga tctgccgccg gactgtgacc
     6541 gccagacgga ttggccgcga cgtgtgcgcg gctacgtcgc taaaggccag ccagtcgtcc
     6601 ctgctcgtca gacagagacg cagagccagc cgaggcgaaa agctctggcc actatgggaa
     6661 gacgtggcgg taaaaaggcc gcagaacgct ggaaagaccc aaacagtgag tacgcccgag
     6721 cacagcgaga aaaactagct aagtccagtc aacgacaagc taggaaagct aaaggaaatc
     6781 gcttgaccat tgcaggttgg tttatgactg ttgagggaga gactggctcg tggccgacaa
     6841 tcaatgaagc tatgtctgaa tttagcgtgt cacgtcagac cgtgaataga gcacttaagg
     6901 tctgcgggca ttgaacttcc acgaggacgc cgaaagcttc ccagtaaatg tgccatctcg
     6961 taggcagaaa acggttcccc cgtagggtct ctctcttggc ctcctttcta ggtcgggctg
     7021 attgctcttg aagctctcta ggggggctca caccataggc agataacgtt ccccaccggc
     7081 tcgcctcgta agcgcacaag gactgctccc aaagatcttc aaagccactg ccgcgactgc
     7141 cttcgcgaag ccttgccccg cggaaatttc ctccaccgag ttcgtgcaca cccctatgcc
     7201 aagcttcttt caccctaaat tcgagagatt ggattcttac cgtggaaatt cttcgcaaaa
     7261 atcgtcccct gatcgccctt gcgacgttgg cgtcggtgcc gctggttgcg cttggcttga
     7321 ccgacttgat cctccggcgt tcagcctgtg ccacagccga caggatggtg accaccattt
     7381 gccccatatc accgtcggta ctgatcccgt cgtcaataaa ccgaaccgct acaccctgag
     7441 catcaaactc ttttatcagt tggatcatgt cggcgtgtcg cggccaagac ggtcgagctt
     7501 cttcaccaga atgacatcac cttcctccac cttcatcctc agcaaatcca gcccttcccg
     7561 atctgttgaa ctgccggatg ccttgtcggt aaagatgcgg ttagctttta cccctgcatc
     7621 tttgagcgct gaggtctgcc tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc
     7681 gccccatcat ccagccagaa agtgagggag ccacggttga tgagagcttt gttgtaggtg
     7741 gaccagttgg tgattttgaa cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga
     7801 tgcgtgatct gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag ccgccgtccc
     7861 gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct gattagaaaa
     7921 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     7981 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     8041 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     8101 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     8161 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     8221 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     8281 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     8341 ggcgcaggaa cactgccagc gcatcagcaa tattttcacc tgaatcagga tattcttcta
     8401 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     8461 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     8521 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     8581 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     8641 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgagc
     8701 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     8761 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     8821 gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca
     8881 cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact
     8941 ggtccaccta caacaaagct ctcatcaacc gtggctccct cactttctgg ctggatgatg
     9001 gggcgattca ggcctggtat gagtcagcaa caccttcttc acgaggcaga cctcagcgct
     9061 agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca
     9121 tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat
     9181 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg
     9241 gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg
     9301 agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa
     9361 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     9421 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
     9481 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt
     9541 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc
     9601 ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc
     9661 actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga
     9721 aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt
     9781 agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga
     9841 gattacgcgc agaccaaaac gatctcaaga agatcatctt attaaggggt ctgacgctca
     9901 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     9961 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    10021 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    10081 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    10141 accatctggc ccca
//
