LOCUS       40924_45468        6697 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pUDI069, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6697)
  AUTHORS   Koopman F, Beekwilder J, Crimi B, Houwelingen A, Hall RD, Bosch D, 
            van Maris AJ, Pronk JT, Daran JM.
  TITLE     De novo production of the flavonoid naringenin in engineered 
            Saccharomyces cerevisiae
  JOURNAL   Microb. Cell Fact. 11 (1), 155 (2012)
  PUBMED    23216753
REFERENCE   2  (bases 1 to 6697)
  AUTHORS   Koopman F, Beekwilder J, Crimi B, van Houwelingen A, Hall RD, Bosch 
            D, Maris AJA., Pronk JT, Daran J-M.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JUL-2012) Platform Green Synthetic Biolog, Delft 2600,
            The Netherlands
REFERENCE   3  (bases 1 to 6697)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6697)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb. 
            Cell Fact."; date: "2012"; volume: "11"; issue: "1"; pages: "155"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-JUL-2012) Platform Green Synthetic Biolog, Delft 2600, The 
            Netherlands"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(148..736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(910..1767)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1768..1872)
                     /label=AmpR promoter
     promoter        2152..2432
                     /label=TRP1 promoter
     CDS             2433..3104
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(3204..3659)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3804..3820
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3827..3845
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3870..4514
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     CDS             4532..6127
                     /codon_start=1
                     /product="tyrosine ammonia lyase"
                     /label=tyrosine ammonia lyase
                     /note="Tal1"
                     /protein_id="AGA15802.1"
                     /translation="MTLQSQTAKDCLALDGALTLVQCEAIATHRSRISVTPALRERCAR
                     AHARLEHAIAEQRHIYGITTGFGPLANRLIGADQGAELQQNLIYHLATGVGPKLSWAEA
                     RALMLARLNSILQGASGASPETIDRIVAVLNAGFAPEVPAQGTVGASGDLTPLAHMVLA
                     LQGRGRMIDPSGRVQEAGAVMDRLCGGPLTLAARDGLALVNGTSAMTAIAALTGVEAAR
                     AIDAALRHSAVLMEVLSGHAEAWHPAFAELRPHPGQLRATERLAQALDGAGRVCRTLTA
                     ARRLTAADLRPEDHPAQDAYSLRVVPQLVGAVWDTLDWHDRVVTCELNSVTDNPIFPEG
                     CAVPALHGGNFMGVHVALASDALNAALVTLAGLVERQIARLTDEKLNKGLPAFLHGGQA
                     GLQSGFMGAQVTATALLAEMRANATPVSVQSLSTNGANQDVVSMGTIAARRARAQLLPL
                     SQIQAILALALAQAMDLLDDPEGQAGWSLTARDLRDRIRAVSPGLRADRPLAGHIEAVA
                     QGLRHPSAAADPPA"
     terminator      6134..6381
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(6404..6422)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6443..6459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6467..6483)
                     /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(6491..6521)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6536..6557)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
       61 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      121 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc ggcccccctg acgagcatca
      181 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      241 gttcccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      301 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
      361 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      421 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      481 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
      541 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
      601 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
      661 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
      721 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
      781 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
      841 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
      901 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
      961 atccatagtt gcctgactgc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1021 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1081 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1141 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1201 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1261 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgaaa
     1321 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1381 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1441 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1501 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1561 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1621 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1681 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     1741 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     1801 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     1861 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     1921 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     1981 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     2041 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     2101 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc aaacgacatt
     2161 actatatata taatatagga agcatttaat agacagcatc gtaatatatg tgtactttgc
     2221 agttatgacg ccagatggca gtagtggaag atattcttta ttgaaaaata gcttgtcacc
     2281 ttacgtacaa tcttgatccg gagcttttct ttttttgccg attaagaatt aattcggtcg
     2341 aaaaaagaaa aggagagggc caagagggag ggcattggtg actattgagc acgtgagtat
     2401 acgtgattaa gcacacaaag gcagcttgga gtatgtctgt tattaatttc acaggtagtt
     2461 ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga ggccgcagaa tgtgctctag
     2521 attccgatgc tgacttgctg ggtattatat gtgtgcccaa tagaaagaga acaattgacc
     2581 cggttattgc aaggaaaatt tcaagtcttg taaaagcata taaaaatagt tcaggcactc
     2641 cgaaatactt ggttggcgtg tttcgtaatc aacctaagga ggatgttttg gctctggtca
     2701 atgattacgg cattgatatc gtccaactgc atggagatga gtcgtggcaa gaataccaag
     2761 agttcctcgg tttgccagtt attaaaagac tcgtatttcc aaaagactgc aacatactac
     2821 tcagtgcagc ttcacagaaa cctcattcgt ttattccctt gtttgattca gaagcaggtg
     2881 ggacaggtga acttttggat tggaactcga tttctgactg ggttggaagg caagagagcc
     2941 ccgaaagctt acattttatg ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc
     3001 ttagattaaa tggcgttatt ggtgttgatg taagcggagg tgtggagaca aatggtgtaa
     3061 aagactctaa caaaatagca aatttcgtca aaaatgctaa gaaataggtt attactgagt
     3121 agtatttatt taagtattgt ttgtgcactt gcctgcggtg tgaaataccg cacagatgcg
     3181 taaggagaaa ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt
     3241 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3301 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3361 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3421 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3481 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3541 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3601 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3661 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     3721 gctattacgc cagctggcga aggggggatg tgctgcaagg cgattaagtt gggtaacgcc
     3781 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact
     3841 atagggcgaa ttggagctct tcgagtttat cattatcaat actcgccatt tcaaagaata
     3901 cgtaaataat taatagtagt gattttccta actttattta gtcaaaaaat tagcctttta
     3961 attctgctgt aacccgtaca tgcccaaaat agggggcggg ttacacagaa tatataacac
     4021 cgaaggtgcc tgggtgaaca ggtttattcc tggcatccac taaatataat ggagcccgct
     4081 ttttaagctg gcatccagaa aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc
     4141 aaccatcagt tcataggtcc attctcttag cgcaactaca gagaacaggg gcacaaacag
     4201 gcaaaaaacg ggcacaacct caatggagtg atgcaacctg cctggagtaa atgatgacac
     4261 aaggcaattg acccacgcat gtatctatct cattttctta caccttctat taccttctgc
     4321 tctctctgat ttggaaaaag ctgaaaaaaa aggttgaaac cagttccctg aaattattcc
     4381 cctacttgac taataagtat ataaagacgg taggtattga ttgtaattct gtaaatctat
     4441 ttcttaaact tcttaaattc tacttttata gttagtcttt tttttagttt taaaacacca
     4501 gaacttagtt tcgacggatt ctagaactag tatgacctta caatcccaaa ctgccaaaga
     4561 ctgcttagcc ttagacggtg ccttgacctt ggttcaatgt gaagcaattg ccacacatag
     4621 atccagaata agtgtcaccc cagctttgag agaaagatgc gctagagcac atgccagatt
     4681 agaacacgct attgcagaac aaagacacat ctatggtata actacaggtt ttggtccttt
     4741 ggctaataga ttaataggtg ccgatcaagg tgctgaattg caacaaaact taatctacca
     4801 tttggctact ggtgttggtc caaaattgtc ttgggccgaa gctagagcat tgatgttggc
     4861 aagattgaac tcaatcttgc aaggtgcatc tggtgcctca cctgaaacaa tcgacagaat
     4921 tgttgctgtc ttaaacgctg gtttcgcacc agaagtccct gcccaaggta ctgtaggtgc
     4981 ttccggtgac ttgacaccat tggcacatat ggttttggcc ttacaaggta gaggtagaat
     5041 gattgatcct agtggtagag ttcaagaagc cggtgctgtc atggacagat tatgtggtgg
     5101 tccattgact ttagctgcaa gagatggttt ggctttagtt aatggtactt ctgccatgac
     5161 agctatcgcc gctttgacag gtgttgaagc agccagagct attgatgctg cattaagaca
     5221 ttccgcagta ttaatggaag ttttgagtgg tcatgcagaa gcctggcacc cagcttttgc
     5281 agaattgaga ccacaccctg gtcaattaag agctaccgaa agattagccc aagctttgga
     5341 tggtgcaggt agagtttgca gaaccttgac tgccgctaga agattgacag cagccgactt
     5401 aagaccagaa gatcatcctg cacaagacgc ctattctttg agagttgtcc cacaattagt
     5461 tggtgctgtc tgggatactt tggactggca cgatagagta gttacctgtg aattgaactc
     5521 agtcactgat aacccaatat ttcctgaagg ttgcgctgta cctgcattac atggtggtaa
     5581 tttcatgggt gtacacgttg cattggcctc cgacgcttta aacgctgcat tagtaacatt
     5641 ggctggttta gttgaaagac aaatcgcaag attgaccgat gaaaagttga ataagggttt
     5701 gccagcattt ttgcatggtg gtcaagcagg tttacaatca ggtttcatgg gtgctcaagt
     5761 tacagctacc gcattgttag cagaaatgag agccaacgct acccctgtct ctgtacaatc
     5821 tttgtcaact aatggtgcta accaagatgt cgtatcaatg ggtactatcg ccgctagaag
     5881 agcaagagcc caattgttgc cattgtctca aatccaagca atcttggctt tagcattggc
     5941 ccaagctatg gacttgttag atgaccctga aggtcaagca ggttggtcct tgacagccag
     6001 agacttaaga gatagaatta gagctgttag tccaggtttg agagctgata gacctttagc
     6061 aggtcatata gaagcagtcg cacaaggttt gagacatcca tccgccgcag cagaccctcc
     6121 agcctaactc gagtcatgta attagttatg tcacgcttac attcacgccc tccccccaca
     6181 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     6241 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     6301 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     6361 cgctcgaagg ctttaatttg cggccggtac ccagcttttg ttccctttag tgagggttaa
     6421 ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6481 caattccaca caacatagga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6541 tgaggtaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6601 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6661 gctcttccgc ttcctcgctc actgactcgc tgcgctc
//
