LOCUS       sequence-95152-e        4598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  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 4598)
  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 4598)
  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-2
FEATURES             Location/Qualifiers
     source          1..4598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..84
                     /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             85..1167
                     /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(104..123)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1399..1404,1405..1421,1422..1428)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1436..1452
                     /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      2595..2666
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2682..2709
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2867..3455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2947..2966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3543..3637)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3668..4462)
                     /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(3779..3798)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4389..4408
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
       61 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
      121 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
      181 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
      241 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
      301 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
      361 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
      421 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
      481 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
      541 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
      601 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
      661 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
      721 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
      781 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
      841 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
      901 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
      961 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     1021 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     1081 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1141 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agcgcgaatt
     1201 gatctggttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca
     1261 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     1321 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     1381 atattctgaa atgagctgtt gacaattaat catccggctc gtataatgtg tggaattgtg
     1441 agcggataac aatttcagaa ttcaaaagat ctaagaagga gatatacata tggcagcagc
     1501 agcagcagca ggtgaagctc gtcgtgttct ggtttacggt ggtcgtggtg cactgggcag
     1561 ccgctgcgtt caggcattcc gtgcgcgtaa ctggtgggtg gcttccgtgg acgttgtcga
     1621 gaacgaagag gcgtccgcat ccattatcgt taaaatgacc gattccttca ccgaacaggc
     1681 agatcaggtt actgccgaag ttggcaaact gctgggtgaa gaaaaagtag atgcgatcct
     1741 gtgtgttgct ggtggctggg ctggtggtaa cgccaaaagc aaatccctgt tcaaaaactg
     1801 tgatctgatg tggaaacagt ctatctggac ctctactatt tctagccatc tggcgactaa
     1861 gcacctgaaa gaaggtggtc tgctgaccct ggccggtgcc aaagcagctc tggatggtac
     1921 gccgggtatg attggttacg gcatggctaa aggcgctgta catcagctgt gccagtccct
     1981 ggcgggtaaa aactctggca tgccaccggg tgctgcggcg atcgcggtgc tgccggtgac
     2041 cctggatacc ccgatgaatc gtaaaagcat gccggaagcg gatttctctt cttggacgcc
     2101 gctggaattt ctggtagaaa cgttccacga ttggattacc ggtaaaaacc gtccgtcttc
     2161 cggtagcctg atccaggtag tgacgaccga aggtcgcacg gaactgacgc cagcctattt
     2221 ttgaactagt aagaaggaga tatacatatg gcaggcaaag cgcaccgtct gtctgcggaa
     2281 gaacgcgatc agctgctgcc gaacctgcgt gcagtgggct ggaacgaact ggaaggtcgc
     2341 gatgctatct tcaaacaatt ccactttaag gatttcaacc gtgcgttcgg cttcatgact
     2401 cgtgttgcac tgcaagcaga aaaactggat catcacccgg aatggttcaa cgtttacaac
     2461 aaagtacata ttaccctgag cacccacgaa tgcgcaggtc tgtctgaacg cgatatcaac
     2521 ctggcgtcct tcatcgaaca ggttgcggtt tccatgacct gaggatccaa actcgagtaa
     2581 ggatctccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     2641 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
     2701 ctttctgcgt ttatacctag ggcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2761 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2821 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2881 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2941 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3001 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3061 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3121 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3181 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3241 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3301 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3361 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3421 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3481 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg actagtgctt
     3541 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     3601 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctctc gaaccccaga
     3661 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     3721 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     3781 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     3841 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     3901 gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg
     3961 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     4021 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     4081 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     4141 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     4201 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     4261 atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     4321 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     4381 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     4441 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcatg
     4501 atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct
     4561 tcccaacctt accagagggc gccccagctg gcaattcc
//
