LOCUS       V003942                10157 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003942
VERSION     V003942
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10157)
  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 10157)
  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 10157)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10157)
  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..10157
                     /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..3429
                     /codon_start=1
                     /product="geranyl diphosphate synthase"
                     /label="geranyl diphosphate synthase"
                     /note="AgGPPS2; codon-optimized for C. glutamicum"
                     /protein_id="AIF29191.1"
                     /translation="MAYSAMATMGYNGMAASCHTLHPTSPLKPFHGASTSLEAFNGEHM
                     GLLRGYSKRKLSSYKNPASRSSNATVAQLLNPPQKGKKAVEFDFNKYMDSKAMTVNEAL
                     NKAIPLRYPQKIYESMRYSLLAGGKRVRPVLCIAACELVGGTEELAIPTACAIEMIHTM
                     SLMHDDLPCIDNDDLRRGKPTNHKIFGEDTAVTAGNALHSYAFEHIAVSTSKTVGADRI
                     LRMVSELGRATGSEGVMGGQMVDIASEGDPSIDLQTLEWIHIHKTAMLLECSVVCGAII
                     GGASEIVIERARRYARCVGLLFQVVDDILDVTKSSDELGKTAGKDLISDKATYPKLMGL
                     EKAKEFSDELLNRAKGELSCFDPVKAAPLLGLADYVAFRQN"
     terminator      3656..3742
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3834..3861
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3880..3971
                     /label="AmpR promoter"
     CDS             complement(7919..8731)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(9326..9871)
                     /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 actccgcaat ggcaaccatg ggctacaacg gcatggcagc atcctgccac
     2341 accctgcacc caacctcccc actgaagcca ttccacggcg catccacctc cctggaagca
     2401 ttcaacggcg aacacatggg cctgctgcgc ggctactcca agcgcaagct gtcctcctac
     2461 aagaacccag catcccgctc ctccaacgca accgtggcac agctgctgaa cccaccacag
     2521 aagggcaaga aggcagtgga gttcgatttc aacaagtaca tggattccaa ggcaatgacc
     2581 gtgaacgaag cactgaacaa ggcaatccca ctgcgctacc cacagaaaat ctacgaatcc
     2641 atgcgctact ccctgctggc aggcggcaag cgcgtgcgcc cagtgctgtg catcgcagca
     2701 tgcgaactgg tgggcggcac cgaagaactg gcaatcccaa ccgcatgcgc aatcgaaatg
     2761 atccacacca tgtccctgat gcacgatgat ctgccatgca tcgataacga tgatctgcgc
     2821 cgcggcaagc caaccaacca caagattttc ggcgaagata ccgcagtgac cgcaggcaac
     2881 gcactgcact cctacgcatt cgaacacatc gcagtgtcca cctccaagac cgtgggcgca
     2941 gatcgcatcc tgcgcatggt gtccgaactg ggccgcgcaa ccggctccga aggcgtgatg
     3001 ggcggccaga tggtggatat cgcatccgaa ggcgatccat ccatcgatct gcagaccctg
     3061 gaatggattc acatccacaa gaccgcaatg ctgctggaat gctccgtggt gtgcggcgca
     3121 atcatcggcg gcgcatccga aatcgtgatc gaacgcgcac gccgctacgc acgctgcgtg
     3181 ggcctgctgt tccaggtggt ggatgatatc ctggatgtga ccaagtcctc cgatgaactg
     3241 ggcaagaccg caggcaagga tctgatctcc gataaggcaa cctacccaaa gctgatgggc
     3301 ctggaaaagg caaaggaatt ttccgatgaa ctgctgaacc gcgcaaaggg cgaactgtcc
     3361 tgcttcgatc cagtgaaggc agcaccactg ctgggcctgg cagattacgt ggcattccgc
     3421 cagaactaag gatccgtcga cctgcagcca agcttctgtt ttggcggatg agagaagatt
     3481 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
     3541 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
     3601 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
     3661 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     3721 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
     3781 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
     3841 catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac
     3901 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3961 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4021 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4081 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4141 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4201 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4261 agaatgactt ggttgagtac tgcggcgtcg ctgatcgccc tggcgacgtt gtgcgggtgg
     4321 cttgtccctg agggcgctgc gacagatagc taaaaatctg ggtcaggatc gccgtagagc
     4381 gcgcgtcgtc gattggaggc ttcccctttg gttgacggtc ttcaatcgct ctacggcgat
     4441 cctgacgctt ttttgttgcg taccgtcgat cgttttattt ctgtcgatcc cgaaaaagtt
     4501 tttgcctttt gtaaaaaact tctcggtcgc cccgcaaatt ttcgattcca gattttttaa
     4561 aaaccaagcc agaaatacga cacaccgttt gcagataatc tgtctttcgg aaaaatcaag
     4621 tgcgatacaa aatttttagc acccctgagc tgcgcaaagt cccgcttcgt gaaaattttc
     4681 gtgccgcgtg attttccgcc aaaaacttta acgaacgttc gttataatgg tgtcatgacc
     4741 ttcacgacga agtactaaaa ttggcccgaa tcatcagcta tggatctctc tgatgtcgcg
     4801 ctggagtccg acgcgctcga tgctgccgtc gatttaaaaa cggtgatcgg atttttccga
     4861 gctctcgata cgacggacgc gccagcatca cgagactggg ccagtgccgc gagcgaccta
     4921 gaaactctcg tggcggatct tgaggagctg gctgacgagc tgcgtgctcg gccagcgcca
     4981 ggaggacgca cagtagtgga ggatgcaatc agttgcgcct actgcggtgg cctgattcct
     5041 ccccggcctg acccgcgagg acggcgcgca aaatattgct cagatgcgtg tcgtgccgca
     5101 gccagccgcg agcgcgccaa caaacgccac gccgaggagc tggaggcggc taggtcgcaa
     5161 atggcgctgg aagtgcgtcc cccgagcgaa attttggcca tggtcgtcac agagctggaa
     5221 gcggcagcga gaattatccg cgatcgtggc gcggtgcccg caggcatgac aaacatcgta
     5281 aatgccgcgt ttcgtgtggc cgtggccgcc caggacgtgt cagcgccgcc accacctgca
     5341 ccgaatcggc agcagcgtcg cgcgtcgaaa aagcgcacag gcggcaagaa gcgataagct
     5401 gcacgaatac ctgaaaaatg ttgaacgccc cgtgagcggt aactcacagg gcgtcggcta
     5461 acccccagtc caaacctggg agaaagcgct caaaaatgac tctagcggat tcacgagaca
     5521 ttgacacacc ggcctggaaa ttttccgctg atctgttcga cacccatccc gagctcgcgc
     5581 tgcgatcacg tggctggacg agcgaagacc gccgcaaatt cctcgctcac ctgggcagag
     5641 aaaatttcca gggcagcaag acccgcgact tcgccagcgc ttggatcaaa gacccggaca
     5701 cgggagaaac acagccgaag ttataccgag ttggttcaaa atcgcttgcc cggtgccagt
     5761 atgttgctct gacgcacgcg cagcacgcag ccgtgcttgt cctggacatt gatgtgccga
     5821 gccaccaggc cggcgggaaa atcgagcacg taaaccccga ggtctacgcg attttggagc
     5881 gctgggcacg cctggaaaaa gcgccagctt ggatcggcgt gaatccactg agcgggaaat
     5941 gccagctcat ctggctcatt gatccggtgt atgccgcagc aggcatgagc agcccgaata
     6001 tgcgcctgct ggctgcaacg accgaggaaa tgacccgcgt tttcggcgct gaccaggctt
     6061 tttcacatag gctgagccgt ggccactgca ctctccgacg atcccagccg taccgctggc
     6121 atgcccagca caatcgcgtg gatcgcctag ctgatcttat ggaggttgct cgcatgatct
     6181 caggcacaga aaaacctaaa aaacgctatg agcaggagtt ttctagcgga cgggcacgta
     6241 tcgaagcggc aagaaaagcc actgcggaag caaaagcact tgccacgctt gaagcaagcc
     6301 tgccgagcgc cgctgaagcg tctggagagc tgatcgacgg cgtccgtgtc ctctggactg
     6361 ctccagggcg tgccgcccgt gatgagacgg cttttcgcca cgctttgact gtgggatacc
     6421 agttaaaagc ggctggtgag cgcctaaaag acaccaaggg tcatcgagcc tacgagcgtg
     6481 cctacaccgt cgctcaggcg gtcggaggag gccgtgagcc tgatctgccg ccggactgtg
     6541 accgccagac ggattggccg cgacgtgtgc gcggctacgt cgctaaaggc cagccagtcg
     6601 tccctgctcg tcagacagag acgcagagcc agccgaggcg aaaagctctg gccactatgg
     6661 gaagacgtgg cggtaaaaag gccgcagaac gctggaaaga cccaaacagt gagtacgccc
     6721 gagcacagcg agaaaaacta gctaagtcca gtcaacgaca agctaggaaa gctaaaggaa
     6781 atcgcttgac cattgcaggt tggtttatga ctgttgaggg agagactggc tcgtggccga
     6841 caatcaatga agctatgtct gaatttagcg tgtcacgtca gaccgtgaat agagcactta
     6901 aggtctgcgg gcattgaact tccacgagga cgccgaaagc ttcccagtaa atgtgccatc
     6961 tcgtaggcag aaaacggttc ccccgtaggg tctctctctt ggcctccttt ctaggtcggg
     7021 ctgattgctc ttgaagctct ctaggggggc tcacaccata ggcagataac gttccccacc
     7081 ggctcgcctc gtaagcgcac aaggactgct cccaaagatc ttcaaagcca ctgccgcgac
     7141 tgccttcgcg aagccttgcc ccgcggaaat ttcctccacc gagttcgtgc acacccctat
     7201 gccaagcttc tttcacccta aattcgagag attggattct taccgtggaa attcttcgca
     7261 aaaatcgtcc cctgatcgcc cttgcgacgt tggcgtcggt gccgctggtt gcgcttggct
     7321 tgaccgactt gatcctccgg cgttcagcct gtgccacagc cgacaggatg gtgaccacca
     7381 tttgccccat atcaccgtcg gtactgatcc cgtcgtcaat aaaccgaacc gctacaccct
     7441 gagcatcaaa ctcttttatc agttggatca tgtcggcgtg tcgcggccaa gacggtcgag
     7501 cttcttcacc agaatgacat caccttcctc caccttcatc ctcagcaaat ccagcccttc
     7561 ccgatctgtt gaactgccgg atgccttgtc ggtaaagatg cggttagctt ttacccctgc
     7621 atctttgagc gctgaggtct gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga
     7681 atcgccccat catccagcca gaaagtgagg gagccacggt tgatgagagc tttgttgtag
     7741 gtggaccagt tggtgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga
     7801 agatgcgtga tctgatcctt caactcagca aaagttcgat ttattcaaca aagccgccgt
     7861 cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga
     7921 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     7981 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     8041 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     8101 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     8161 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     8221 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     8281 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     8341 accggcgcag gaacactgcc agcgcatcag caatattttc acctgaatca ggatattctt
     8401 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     8461 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     8521 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     8581 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     8641 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     8701 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     8761 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     8821 tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga
     8881 tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca
     8941 actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg
     9001 atggggcgat tcaggcctgg tatgagtcag caacaccttc ttcacgaggc agacctcagc
     9061 gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct
     9121 tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata
     9181 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     9241 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     9301 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     9361 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     9421 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     9481 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     9541 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     9601 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     9661 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     9721 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     9781 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     9841 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaagg ggtctgacgc
     9901 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     9961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    10021 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    10081 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    10141 cttaccatct ggcccca
//
