LOCUS       V002346                 8101 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002346
VERSION     V002346
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8101)
  AUTHORS   Kozak BU, van Rossum HM, Benjamin KR, Wu L, Daran JM, Pronk JT, van
            Maris AJ.
  TITLE     Replacement of the Saccharomyces cerevisiae acetyl-CoA synthetases
            by alternative pathways for cytosolic acetyl-CoA synthesis
  JOURNAL   Metab. Eng. (2013) In press
   PUBMED   24269999
REFERENCE   2  (bases 1 to 8101)
  AUTHORS   Kozak BU, van Rossum HM, Benjamin KR, Wu L, Daran J-M.G., Pronk JT,
            van Maris AJA.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAY-2013) Biotechnology, Delft University of
            Technology, Julianalaan 67, Delft 2628BC, The Netherlands
REFERENCE   3  (bases 1 to 8101)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8101)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
            (2013) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (31-MAY-2013) Biotechnology, Delft University of Technology,
            Julianalaan 67, Delft 2628BC, The Netherlands"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(945..1802)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(1803..1907)
                     /label="AmpR promoter"
     rep_origin      complement(1934..3276)
                     /direction=LEFT
                     /label="2u ori"
                     /note="yeast 2u plasmid origin of replication"
     promoter        3540..3760
                     /label="URA3 promoter"
     CDS             3761..4561
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     rep_origin      complement(4695..5150)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     5295..5311
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        5321..5339
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(5358..5605)
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     primer_bind     5608..5624
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    5657..5681
                     /label="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5700..7100)
                     /gene="eutE"
                     /label="Acetaldehyde dehydrogenase (acetylating) EutE"
                     /note="Acetaldehyde dehydrogenase (acetylating) EutE from
                     Escherichia coli (strain K12). Accession#: P77445"
     protein_bind    complement(7116..7140)
                     /label="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7148..7164)
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        complement(7170..7814)
                     /label="GAP promoter"
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     promoter        complement(7843..7861)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7882..7898)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7906..7922)
                     /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)."
     promoter        complement(7930..7960)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7975..7996)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     1981 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     2041 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     2101 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2161 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     2221 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     2281 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     2341 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     2401 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     2461 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     2521 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     2581 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     2641 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     2701 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     2761 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
     2821 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg
     2881 ttttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa
     2941 cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg cgcacataca
     3001 gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg tatatatata tacatgagaa
     3061 gaacggcata gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat ttatgtagga
     3121 tgaaaggtag tctagtacct cctgtgatat tatcccattc catgcggggt atcgtatgct
     3181 tccttcagca ctacccttta gctgttctat atgctgccac tcctcaattg gattagtctc
     3241 atccttcaat gctatcattt cctttgatat tggatcatac taagaaacca ttattatcat
     3301 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     3361 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     3421 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     3481 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata ccacagcttt
     3541 tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc tttgaaattt
     3601 ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt
     3661 ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca
     3721 actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag ctacatataa
     3781 ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga
     3841 aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt
     3901 agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga
     3961 tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt
     4021 actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc
     4081 gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc
     4141 aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct
     4201 tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg
     4261 tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga
     4321 catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga
     4381 tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg tctctacagg
     4441 atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga
     4501 gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa
     4561 ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat
     4621 ttaattatat cagttattac cctatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4681 aataccgcat caggaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     4741 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4801 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4861 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4921 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4981 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     5041 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     5101 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cgcgccattc
     5161 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5221 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5281 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     5341 cgaattgggt accggccgca aattaaagcc ttcgagcgtc ccaaaacctt ctcaagcaag
     5401 gttttcagta taatgttaca tgcgtacacg cgtctgtaca gaaaaaaaag aaaaatttga
     5461 aatataaata acgttcttaa tactaacata actataaaaa aataaatagg gacctagact
     5521 tcaggttgtc taactccttc cttttcggtt agagcggatg tggggggagg gcgtgaatgt
     5581 aagcgtgaca taactaatta catgactcga ggtcgacggt atcgataagc ttgatatcga
     5641 attcctgcag cccatcacca ctttgtacaa gaaagctggg tgtcgactag gaattcttaa
     5701 acgattctga aagcgtcaac caaaacacat cttctcaatc taacgaaagt tctagcagaa
     5761 gtaacacctt caccagttgg agtagtgata gtcatagtag tccaaccttc accacccaaa
     5821 cccaaaccag cgatacatgg accgttctta acgaagatag aagtgtcgat agcgttagcc
     5881 atttggttca tgttttcgat gtttctagag tgcatagcag cagtgtggtg acaaccacct
     5941 tccaacttaa cagccaaagc gatagcgtca gcaacgttag caactctaac aactggcaaa
     6001 actggcatca tcaattcagt aacagcgaat gggtgttcag cagtagtttc aacgaacaac
     6061 aatctagttt cttgtggaac gttcaaaccg atagcagcag cgatcttagc agcgtctcta
     6121 ccaacccagt ctctagaaac agtaccctta cctctttcgt cgatgttctt caacaaaact
     6181 ggttgcaatt gttgagcttg ttcagcagtc aacttaacag cgtgttgacc ttccatcaat
     6241 ctcatcaatt cgtcagcaac agagtcaaca acgatcaaaa ccttttcgtc agcacagatg
     6301 atgttgttgt cgaaagaagc acccttaacg atagattgag cagctctagc caagtcagca
     6361 gtttcgtcaa caacaactgg tgggttacca gcaccagcag cgatcaatct cttgttagtg
     6421 tgctttctag cagcttcaac aacagcttca ccaccagtaa caaccaacaa accgatacct
     6481 gggaacttga acaatctttg agcagtttcg atgtctgggt tagcaacagt aaccaacaag
     6541 ttttctggac caccagcagc aacgatagct tggttcaaca aagtgatagc tctttgagaa
     6601 accttcttag cagctgggtg tggagcgaag ataacagagt taccagcagc gatcaaagag
     6661 atagcgttgt tgatcaaagt agcagctggg ttagtagatg gagtaacaga agcaacaaca
     6721 ccccatggag cgttttcgat caaagtcaaa ccgttgtcac cagtcaaaac ttgtggagac
     6781 aaacattcaa cacctggagt acctctagct tgagcaacgt tcttagcgaa cttgtcttca
     6841 actctaccca taccagtttc agaaacagcc aattcagcca agtctctagc gtgcttttca
     6901 ccagcttctc tgatagcagc gatagccaat tgtctcatag caacagactt caaaccttgt
     6961 tgagcaacct tagcagcagc aacagcgtcg tccaaagaag cgaaaacacc catttcgtga
     7021 acagcagctg gtggagtgtc agaagattgc atcttcaaca aaacagcctt aacaacttgt
     7081 tcgatgtctt gttggttcat ggatcctatc tgcagagcct gcttttttgt acaaacttgt
     7141 gatgggggat ccactagttc tagaatccgt cgaaactaag ttctggtgtt ttaaaactaa
     7201 aaaaaagact aactataaaa gtagaattta agaagtttaa gaaatagatt tacagaatta
     7261 caatcaatac ctaccgtctt tatatactta ttagtcaagt aggggaataa tttcagggaa
     7321 ctggtttcaa cctttttttt cagctttttc caaatcagag agagcagaag gtaatagaag
     7381 gtgtaagaaa atgagataga tacatgcgtg ggtcaattgc cttgtgtcat catttactcc
     7441 aggcaggttg catcactcca ttgaggttgt gcccgttttt tgcctgtttg tgcccctgtt
     7501 ctctgtagtt gcgctaagag aatggaccta tgaactgatg gttggtgaag aaaacaatat
     7561 tttggtgctg ggattctttt tttttctgga tgccagctta aaaagcgggc tccattatat
     7621 ttagtggatg ccaggaataa acctgttcac ccaggcacct tcggtgttat atattctgtg
     7681 taacccgccc cctattttgg gcatgtacgg gttacagcag aattaaaagg ctaatttttt
     7741 gactaaataa agttaggaaa atcactacta ttaattattt acgtattctt tgaaatggcg
     7801 agtattgata atgataaact cgaagagctc cagcttttgt tccctttagt gagggttaat
     7861 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     7921 aattccacac aacataggag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     7981 gaggtaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     8041 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     8101 c
//
