LOCUS       sequence-129984-        6117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tyrosine hydroxylase from mouse, dihydropterine reductase from 
            human, pterin-4a-carbinolamine dehydratase from human.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  AUTHORS   Satoh Y, Tajima K, Munekata M, Keasling JD, Lee TS
  TITLE     Engineering of L-tyrosine oxidation in Escherichia coli and 
            microbial production of hydroxytyrosol.
  JOURNAL   Metab Eng. 2012 Nov;14(6):603-10. doi: 10.1016/j.ymben.2012.08.002. 
            Epub 2012 Aug 29.
  PUBMED    22948011
REFERENCE   2  (bases 1 to 6117)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ymben.2012.08.002"; journalName: "Metab Eng"; date: 
            "2012-11"; volume: "14"; issue: "6"; pages: "603-10"
COMMENT     Sequence Label: pBbE1k-3
FEATURES             Location/Qualifiers
     source          1..6117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(26..820)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(137..156)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     747..766
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        963..1040
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             1041..2123
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1060..1079)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2355..2360,2361..2377,2378..2384)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2392..2408
                     /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)."
     terminator      5070..5141
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5157..5184
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(5342..5930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5422..5441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6018..6112)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 gagctctcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
       61 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
      121 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
      181 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
      241 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga
      301 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
      361 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
      421 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
      481 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
      541 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
      601 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
      661 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
      721 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
      781 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
      841 tctcttgatc agatcatgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
      901 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccgacg
      961 tcgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     1021 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     1081 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     1141 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     1201 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     1261 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     1321 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     1381 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     1441 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     1501 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     1561 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     1621 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     1681 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     1741 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     1801 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     1861 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     1921 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1981 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     2041 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2101 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagcg cgaattgatc
     2161 tggtttgaca gcttatcatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca
     2221 tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag
     2281 gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat
     2341 tctgaaatga gctgttgaca attaatcatc cggctcgtat aatgtgtgga attgtgagcg
     2401 gataacaatt tcagaattca aagatctaag aaggagatat acatatgccg accccgagcg
     2461 cgagctcccc gcagcctaaa ggcttccgtc gcgccgtttc cgaacaggat acgaaacagg
     2521 cagaagcggt aaccagccca cgtttcatcg gtcgccgcca gtccctgatt gaggatgctc
     2581 gtaaggaacg tgaagctgcc gcagcggccg cagcggcagc ggttgctagc gctgaaccgg
     2641 gtaatccgct ggaagccgtg gtgttcgaag aacgtgacgg caacgccgtg ctgaacctgc
     2701 tgttctccct gcgtggcacc aagccgtctt ccctgtctcg cgccctgaaa gtttttgaaa
     2761 ctttcgaagc gaaaatccac catctggaaa cccgtcctgc tcagcgcccg ctggcgggca
     2821 gcccacacct ggaatacttt gttcgtttcg aagtgccgtc tggcgatctg gccgcgctgc
     2881 tgtctagcgt tcgtcgtgtg tctgatgacg tgcgttctgc gcgcgaagac aaagtgccgt
     2941 ggttcccgcg caaggtgtct gaactggaca agtgccacca tctggttacc aaattcgatc
     3001 cggacctgga tctggaccat ccgggcttct ccgaccaagc gtaccgtcaa cgccgcaagc
     3061 tgatcgcaga gatcgcgttt cagtataaac agggtgagcc gatcccgcat gtggagtata
     3121 ctaaagaaga gatcgcgact tggaaagaag tgtatgcaac cctgaaaggc ctgtatgcca
     3181 ctcatgcttg ccgcgaacac ctggaagctt tccagctgct ggaacgttac tgtggctacc
     3241 gtgaggattc tattccgcaa ctggaggacg tgtcccactt cctgaaagaa cgtaccggtt
     3301 tccagctgcg tccggtagct ggtctgctgt ccgcgcgtga tttcctggct tccctggcat
     3361 tccgtgtttt ccagtgcacg cagtacatcc gccacgcctc ctccccgatg cattccccgg
     3421 aaccggactg ttgccatgaa ctgctgggtc atgttcctat gctggctgat cgtacttttg
     3481 ctcaattcag ccaggacatt ggcctggcgt ctctgggtgc ctccgatgaa gagatcgaaa
     3541 aactgagcac cgtgtactgg ttcaccgttg agttcggcct gtgcaaacag aacggtgaac
     3601 tgaaagcata tggcgccggt ctgctgtcca gctacggtga actgctgcac tctctgtctg
     3661 aagagccgga agtgcgtgcg ttcgatcctg atactgctgc ggtgcagccg taccaggacc
     3721 agacctacca gccagtttat ttcgtgtctg agtctttctc cgacgctaaa gataaactgc
     3781 gcaactacgc ttcccgtatc cagcgtccgt ttagcgtaaa atttgatccg tataccctgg
     3841 caattgacgt tctggacagc ccgcacacca tccgtcgtag cctggaaggc gttcaggatg
     3901 aactgcatac gctgacccag gcactgagcg ctatcagctg aggatctaag aaggagatat
     3961 acatatggca gcagcagcag cagcaggtga agctcgtcgt gttctggttt acggtggtcg
     4021 tggtgcactg ggcagccgct gcgttcaggc attccgtgcg cgtaactggt gggtggcttc
     4081 cgtggacgtt gtcgagaacg aagaggcgtc cgcatccatt atcgttaaaa tgaccgattc
     4141 cttcaccgaa caggcagatc aggttactgc cgaagttggc aaactgctgg gtgaagaaaa
     4201 agtagatgcg atcctgtgtg ttgctggtgg ctgggctggt ggtaacgcca aaagcaaatc
     4261 cctgttcaaa aactgtgatc tgatgtggaa acagtctatc tggacctcta ctatttctag
     4321 ccatctggcg actaagcacc tgaaagaagg tggtctgctg accctggccg gtgccaaagc
     4381 agctctggat ggtacgccgg gtatgattgg ttacggcatg gctaaaggcg ctgtacatca
     4441 gctgtgccag tccctggcgg gtaaaaactc tggcatgcca ccgggtgctg cggcgatcgc
     4501 ggtgctgccg gtgaccctgg ataccccgat gaatcgtaaa agcatgccgg aagcggattt
     4561 ctcttcttgg acgccgctgg aatttctggt agaaacgttc cacgattgga ttaccggtaa
     4621 aaaccgtccg tcttccggta gcctgatcca ggtagtgacg accgaaggtc gcacggaact
     4681 gacgccagcc tatttttgaa ctagtaagaa ggagatatac atatggcagg caaagcgcac
     4741 cgtctgtctg cggaagaacg cgatcagctg ctgccgaacc tgcgtgcagt gggctggaac
     4801 gaactggaag gtcgcgatgc tatcttcaaa caattccact ttaaggattt caaccgtgcg
     4861 ttcggcttca tgactcgtgt tgcactgcaa gcagaaaaac tggatcatca cccggaatgg
     4921 ttcaacgttt acaacaaagt acatattacc ctgagcaccc acgaatgcgc aggtctgtct
     4981 gaacgcgata tcaacctggc gtccttcatc gaacaggttg cggtttccat gacctgagga
     5041 tccaaactcg agtaaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact
     5101 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     5161 caccttcggg tgggcctttc tgcgtttata cctagggcgt tcggctgcgg cgagcggtat
     5221 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5281 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5341 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5401 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5461 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5521 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5581 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5641 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5701 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5761 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5821 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5881 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5941 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6001 tcatgactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
     6061 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagc
//
