LOCUS       V003943                10163 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003943
VERSION     V003943
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10163)
  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 10163)
  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 10163)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10163)
  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..10163
                     /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..2256
                     /gene="PT1"
                     /label="(-)-alpha-pinene synthase, chloroplastic"
                     /note="(-)-alpha-pinene synthase, chloroplastic from Pinus
                     taeda. Accession#: Q84KL6"
     RBS             2266..2277
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     CDS             2284..3435
                     /codon_start=1
                     /product="geranyl diphosphate synthase"
                     /label="geranyl diphosphate synthase"
                     /note="prenyltransferase AgGPPS1; codon-optimized for C.
                     glutamicum"
                     /protein_id="AIF29193.1"
                     /translation="MAYSCMAASCHGLHFMNIASQECNLKRGIIPSKRLHGISSSLWAS
                     NGFQGHLERDLSAYRHLVSSSRCLNTIAMLSNLSEQAKEKATEFDFKEYLHSKAISVNE
                     ALERAVPLRYPEKIHEAMRYSLLAGGKRIRPILTIAACELVGGSEELAMPTACAMEMIH
                     TMSLIHDDLPSMDNDDLRRGKLTNHKVFGEGTAVLAGDALLSFAFEHIAVSTRKTVASH
                     RVLRVVSELGKAIGSQGVAGGQVADITSEGDPSVGLETLEWIHIHKTAVLLECAVVSGA
                     IIGGASENEIERTGRYARCVGLLFQVVDDILDVTRSSEELGKTAGKDLVSDKATYPKLM
                     GLEKAKEFADELLDRAKEELSCFNPAKAAPLLGLADYIALRQN"
     terminator      3662..3748
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3840..3867
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3886..3977
                     /label="AmpR promoter"
     CDS             complement(7925..8737)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(9332..9877)
                     /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 tgtccccagt ctccgtcatc tccctgccat ccgacctgtg cctgccaacc
      421 tccttcatcg accgctccgg ccgcgaactg atcccactgc acatcaccat cccaaacgtc
      481 gcaatgcgcc gccagggcaa gctgatgacc cgcgcatcca tgtccatgaa cctgcgcacc
      541 gcagtctccg acgacgcagt catccgccgc cgcggcgact tccactccaa cctgtgggac
      601 gacgacctga tccagtccct gtcctcccca tacggcgaac catcctaccg cgaacgcgca
      661 gaacgcctga tcggcgaagt caagaactcc ttcaactcca tgtccaacga agacggcgaa
      721 tccatcaccc cactggacga cctgatccag cgcctgtgga tggtcgactc cgtcgaacgc
      781 ctgggcatcg accgccactt caagaaggaa atcaagtccg cactggacca cgtctaccgc
      841 tactggtccg aaaagggcat cggctgcggc cgcgaatccg tcgtcaccga cctgaactcc
      901 accgcactgg gcctgcgcac cctgcgcctg cacggctacg acgtctccgc agacgtcctg
      961 aaccacttca agaaccagtc cggccagttc gcatgcaccc tgaagcagac cgaagaccag
     1021 atccgcaccg tcctgaacct gtaccgcgca tccctgatcg cattcccagg cgaaaaggtc
     1081 atggacgaag cagaatcctt ctccgcaaag tacctgaagg aagcactcca gaagatccca
     1141 gtctcctcct tctcccgcga aatcggcgac gtcctggaat acggctggca cacctacctg
     1201 ccacgcctgg aagcacgcaa ctacatcgac gtcttcggcc aggacaccga aaactccaag
     1261 tcctacatga agaccgaaaa gctgctggaa ctggcaaagc tggagttcaa catcttccac
     1321 gcactccaga agcgcgaact ggaatacctg gtccgctggt ggaagggctc cggctcccca
     1381 cagatgacct tctgccgcca ccgccacgtc gaatactaca ccctggcatc ctgcatcgca
     1441 ttcgaaccac agcactccgg cttccgcctg ggcttcgcaa aggcatgcca catcatcacc
     1501 gtcctggacg acatgtacga caccttcggc accctggacg aactggaact gttcacctcc
     1561 gcaatcaagc gctgggaccc atccgcaacc gaatgcctgc cagaatacat gaagggcgtc
     1621 tacatgatcg tctacaacac cgtcaacgaa atgtcccagg aagcagacaa ggcacagggc
     1681 cgcgacaccc tgaactactg ccgccaggca tgggaagaat acatcgacgc atacatgcag
     1741 gaagcaaagt ggatcgcatc cggcgaagtc ccaaccttcg aagaatacta cgaaaacggc
     1801 aaggtttcct ccggccaccg cgtctccgca ctccagccaa tcctgaccac cgacatccca
     1861 ttcccagaac acgtcctgaa ggaagtcgac atcccatccc agctgaacga cctggcatcc
     1921 gcaatcctgc gcctgcgcgg cgacacccgc tgctaccagg cagaccgcgc acgcggcgaa
     1981 gaagcatcct gcatctcctg ctacatgaag gacaacccag gcaccaccga agaagacgca
     2041 ctgaaccacc tgaacgcaat gatctccgac gtcatcaagg gcctgaactg ggaactgctg
     2101 aagccaaact cctccgtccc aatctccgca aagaagcacg cattcgacat ctcccgcgca
     2161 ttccactgcg gctacaagta ccgcgacggc tactccgtcg caaacatcga aaccaagtcc
     2221 ctggtcaagc gcaccgtcat cgacccagtc accctgtaag gatctaaaga ggagaaatac
     2281 tagatggcat actcctgcat ggcagcatcc tgccacggcc tgcacttcat gaacatcgca
     2341 tcccaggaat gcaacctgaa gcgcggcatc atcccatcca agcgcctgca cggcatctcc
     2401 tcctccctgt gggcatccaa cggcttccag ggccacctgg aacgcgacct gtccgcatac
     2461 cgccacctgg tttcctcctc ccgctgcctg aacaccatcg caatgctgtc caacctgtcc
     2521 gaacaggcaa aggaaaaggc aaccgagttc gacttcaagg aatacctgca ctccaaggca
     2581 atctccgtca acgaagcact ggaacgcgca gtcccactgc gctacccaga aaagatccac
     2641 gaagcaatgc gctactccct gctggcaggc ggcaagcgca tccgcccaat cctgaccatc
     2701 gcagcatgcg aactggtcgg cggctccgaa gaactggcaa tgccaaccgc atgcgcaatg
     2761 gaaatgatcc acaccatgtc cctgatccac gacgacctgc catccatgga caacgacgac
     2821 ctgcgccgcg gcaagctgac caaccacaag gtcttcggcg aaggcaccgc agtcctggca
     2881 ggcgacgcac tgctgtcctt cgcattcgaa cacatcgcag tctccacccg caagaccgtc
     2941 gcatcccacc gcgtcctgcg cgtcgtctcc gaactgggca aggcaatcgg ctcccagggc
     3001 gtcgcaggcg gccaggtcgc agacatcacc tccgaaggcg acccatccgt cggcctggaa
     3061 accctggaat ggattcacat ccacaagacc gcagtcctgc tggaatgcgc agtcgtctcc
     3121 ggcgcaatca tcggcggcgc atccgaaaac gaaatcgaac gcaccggccg ctacgcacgc
     3181 tgcgtcggcc tgctgttcca ggtcgtcgac gacatcctgg acgtcacccg ctcctccgaa
     3241 gaactgggca agaccgcagg caaggacctg gtttccgaca aggcaaccta cccaaagctg
     3301 atgggcctgg aaaaggcaaa ggagttcgca gacgaactgc tggaccgcgc aaaggaagaa
     3361 ctgtcctgct tcaacccagc aaaggcagca ccactgctgg gcctggcaga ctacatcgca
     3421 ctgcgccaga actaaggatc cgtcgacctg cagccaagct tctgttttgg cggatgagag
     3481 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     3541 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     3601 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     3661 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     3721 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     3781 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     3841 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     3901 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3961 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4021 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4081 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4141 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4201 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     4261 attctcagaa tgacttggtt gagtactgcg gcgtcgctga tcgccctggc gacgttgtgc
     4321 gggtggcttg tccctgaggg cgctgcgaca gatagctaaa aatctgggtc aggatcgccg
     4381 tagagcgcgc gtcgtcgatt ggaggcttcc cctttggttg acggtcttca atcgctctac
     4441 ggcgatcctg acgctttttt gttgcgtacc gtcgatcgtt ttatttctgt cgatcccgaa
     4501 aaagtttttg ccttttgtaa aaaacttctc ggtcgccccg caaattttcg attccagatt
     4561 ttttaaaaac caagccagaa atacgacaca ccgtttgcag ataatctgtc tttcggaaaa
     4621 atcaagtgcg atacaaaatt tttagcaccc ctgagctgcg caaagtcccg cttcgtgaaa
     4681 attttcgtgc cgcgtgattt tccgccaaaa actttaacga acgttcgtta taatggtgtc
     4741 atgaccttca cgacgaagta ctaaaattgg cccgaatcat cagctatgga tctctctgat
     4801 gtcgcgctgg agtccgacgc gctcgatgct gccgtcgatt taaaaacggt gatcggattt
     4861 ttccgagctc tcgatacgac ggacgcgcca gcatcacgag actgggccag tgccgcgagc
     4921 gacctagaaa ctctcgtggc ggatcttgag gagctggctg acgagctgcg tgctcggcca
     4981 gcgccaggag gacgcacagt agtggaggat gcaatcagtt gcgcctactg cggtggcctg
     5041 attcctcccc ggcctgaccc gcgaggacgg cgcgcaaaat attgctcaga tgcgtgtcgt
     5101 gccgcagcca gccgcgagcg cgccaacaaa cgccacgccg aggagctgga ggcggctagg
     5161 tcgcaaatgg cgctggaagt gcgtcccccg agcgaaattt tggccatggt cgtcacagag
     5221 ctggaagcgg cagcgagaat tatccgcgat cgtggcgcgg tgcccgcagg catgacaaac
     5281 atcgtaaatg ccgcgtttcg tgtggccgtg gccgcccagg acgtgtcagc gccgccacca
     5341 cctgcaccga atcggcagca gcgtcgcgcg tcgaaaaagc gcacaggcgg caagaagcga
     5401 taagctgcac gaatacctga aaaatgttga acgccccgtg agcggtaact cacagggcgt
     5461 cggctaaccc ccagtccaaa cctgggagaa agcgctcaaa aatgactcta gcggattcac
     5521 gagacattga cacaccggcc tggaaatttt ccgctgatct gttcgacacc catcccgagc
     5581 tcgcgctgcg atcacgtggc tggacgagcg aagaccgccg caaattcctc gctcacctgg
     5641 gcagagaaaa tttccagggc agcaagaccc gcgacttcgc cagcgcttgg atcaaagacc
     5701 cggacacggg agaaacacag ccgaagttat accgagttgg ttcaaaatcg cttgcccggt
     5761 gccagtatgt tgctctgacg cacgcgcagc acgcagccgt gcttgtcctg gacattgatg
     5821 tgccgagcca ccaggccggc gggaaaatcg agcacgtaaa ccccgaggtc tacgcgattt
     5881 tggagcgctg ggcacgcctg gaaaaagcgc cagcttggat cggcgtgaat ccactgagcg
     5941 ggaaatgcca gctcatctgg ctcattgatc cggtgtatgc cgcagcaggc atgagcagcc
     6001 cgaatatgcg cctgctggct gcaacgaccg aggaaatgac ccgcgttttc ggcgctgacc
     6061 aggctttttc acataggctg agccgtggcc actgcactct ccgacgatcc cagccgtacc
     6121 gctggcatgc ccagcacaat cgcgtggatc gcctagctga tcttatggag gttgctcgca
     6181 tgatctcagg cacagaaaaa cctaaaaaac gctatgagca ggagttttct agcggacggg
     6241 cacgtatcga agcggcaaga aaagccactg cggaagcaaa agcacttgcc acgcttgaag
     6301 caagcctgcc gagcgccgct gaagcgtctg gagagctgat cgacggcgtc cgtgtcctct
     6361 ggactgctcc agggcgtgcc gcccgtgatg agacggcttt tcgccacgct ttgactgtgg
     6421 gataccagtt aaaagcggct ggtgagcgcc taaaagacac caagggtcat cgagcctacg
     6481 agcgtgccta caccgtcgct caggcggtcg gaggaggccg tgagcctgat ctgccgccgg
     6541 actgtgaccg ccagacggat tggccgcgac gtgtgcgcgg ctacgtcgct aaaggccagc
     6601 cagtcgtccc tgctcgtcag acagagacgc agagccagcc gaggcgaaaa gctctggcca
     6661 ctatgggaag acgtggcggt aaaaaggccg cagaacgctg gaaagaccca aacagtgagt
     6721 acgcccgagc acagcgagaa aaactagcta agtccagtca acgacaagct aggaaagcta
     6781 aaggaaatcg cttgaccatt gcaggttggt ttatgactgt tgagggagag actggctcgt
     6841 ggccgacaat caatgaagct atgtctgaat ttagcgtgtc acgtcagacc gtgaatagag
     6901 cacttaaggt ctgcgggcat tgaacttcca cgaggacgcc gaaagcttcc cagtaaatgt
     6961 gccatctcgt aggcagaaaa cggttccccc gtagggtctc tctcttggcc tcctttctag
     7021 gtcgggctga ttgctcttga agctctctag gggggctcac accataggca gataacgttc
     7081 cccaccggct cgcctcgtaa gcgcacaagg actgctccca aagatcttca aagccactgc
     7141 cgcgactgcc ttcgcgaagc cttgccccgc ggaaatttcc tccaccgagt tcgtgcacac
     7201 ccctatgcca agcttctttc accctaaatt cgagagattg gattcttacc gtggaaattc
     7261 ttcgcaaaaa tcgtcccctg atcgcccttg cgacgttggc gtcggtgccg ctggttgcgc
     7321 ttggcttgac cgacttgatc ctccggcgtt cagcctgtgc cacagccgac aggatggtga
     7381 ccaccatttg ccccatatca ccgtcggtac tgatcccgtc gtcaataaac cgaaccgcta
     7441 caccctgagc atcaaactct tttatcagtt ggatcatgtc ggcgtgtcgc ggccaagacg
     7501 gtcgagcttc ttcaccagaa tgacatcacc ttcctccacc ttcatcctca gcaaatccag
     7561 cccttcccga tctgttgaac tgccggatgc cttgtcggta aagatgcggt tagcttttac
     7621 ccctgcatct ttgagcgctg aggtctgcct cgtgaagaag gtgttgctga ctcataccag
     7681 gcctgaatcg ccccatcatc cagccagaaa gtgagggagc cacggttgat gagagctttg
     7741 ttgtaggtgg accagttggt gattttgaac ttttgctttg ccacggaacg gtctgcgttg
     7801 tcgggaagat gcgtgatctg atccttcaac tcagcaaaag ttcgatttat tcaacaaagc
     7861 cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg
     7921 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     7981 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     8041 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     8101 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     8161 ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc
     8221 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     8281 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     8341 aatgcaaccg gcgcaggaac actgccagcg catcagcaat attttcacct gaatcaggat
     8401 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     8461 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     8521 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     8581 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     8641 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     8701 gcctcgagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     8761 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     8821 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     8881 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
     8941 tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc
     9001 tggatgatgg ggcgattcag gcctggtatg agtcagcaac accttcttca cgaggcagac
     9061 ctcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     9121 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     9181 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     9241 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     9301 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     9361 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     9421 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
     9481 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
     9541 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
     9601 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
     9661 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
     9721 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
     9781 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
     9841 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaaggggtc
     9901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     9961 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
    10021 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
    10081 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
    10141 ggagggctta ccatctggcc cca
//
