LOCUS       sequence-96597-e        5083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast D-Amino acid oxidase from Rhodotorula gracilis, optimized for 
            mammalian expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5083)
  AUTHORS   Matlashov ME, Belousov VV, Enikolopov G
  TITLE     How much H(2)O(2) is produced by recombinant D-amino acid oxidase in
            mammalian cells?
  JOURNAL   Antioxid Redox Signal. 2014 Mar 1;20(7):1039-44. doi: 
            10.1089/ars.2013.5618. Epub 2013 Oct 23.
  PUBMED    24020354
REFERENCE   2  (bases 1 to 5083)
  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: 
            "10.1089/ars.2013.5618"; journalName: "Antioxid Redox Signal"; date:
            "2014-03-1- 1"; volume: "20"; issue: "7"; pages: "1039-44"
COMMENT     Sequence Label: pCMV-DAAO
FEATURES             Location/Qualifiers
     source          1..5083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1871..1992
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1908..1927)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1962..1981
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1999..2454)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2136..2157)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2348..2367
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2481..2585
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2587..2944
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2587..2778
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2610..2630)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2795..2930
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2979..3773
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2979..3773
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3033..3052)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3643..3662
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3961..3980)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4005..4052
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4381..4969
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4870..4889
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcaccg
      601 ccatgcacag ccagaagagg gtggtcgtgc tgggctccgg agtcatcggg ctctctagcg
      661 ccctgattct cgctagaaaa ggctactccg tgcatatcct ggccagagac ttgcctgagg
      721 atgtcagctc ccagaccttc gcaagcccct gggccggagc taactggaca ccatttatga
      781 ccctgacaga cggacccaga caagccaagt gggaagagtc caccttcaag aaatgggtgg
      841 agctggttcc cacaggccac gctatgtggc tgaaagggac cagaaggttt gcccagaatg
      901 aggatggcct cctgggacat tggtacaagg acatcacccc taactatagg ccacttccaa
      961 gctctgaatg tccacctgga gccattggag tgacatacga caccctgagc gtccacgctc
     1021 ccaagtattg ccaatacctt gccagagagc tgcagaaact tggggctacc ttcgaaagga
     1081 gaacagtgac tagcctggag caagccttcg atggagctga cctcgtggtc aacgccaccg
     1141 ggctgggagc taagtccatt gccggcatcg atgaccaggc tgccgagccc attaggggac
     1201 agaccgtgct tgtcaaatct ccctgtaaga ggtgcacaat ggatagctcc gaccctgctt
     1261 ctcccgctta tatcattcct agaccaggag gagaagtgat ctgtggcgga acctacggag
     1321 tcggcgattg ggacctgagc gtgaatcccg aaacagtcca aaggattctc aagcactgcc
     1381 tgagacttga ccctaccatt agctccgatg gaactatcga gggcattgaa gtgctgagac
     1441 ataacgttgg cctgagaccc gccaggagag gaggacctag ggtggaggct gaaagaatcg
     1501 tcttgcccct ggaccggacc aagagccctc tctctctggg aagagggtcc gcacgggctg
     1561 caaaagagaa ggaagtgacc cttgtccacg cctacggctt tagtagcgca gggtatcagc
     1621 aatcctgggg agccgctgag gacgtggctc agctggtcga tgaagccttc caaagatacc
     1681 acggcgcagc tagggagagc tgaaagcttc gaattctgca gtcgacggta ccgcgggccc
     1741 gggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     1801 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     1861 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1921 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1981 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2041 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2101 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2161 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2221 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2281 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2341 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2401 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     2461 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2521 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2581 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2641 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2701 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2761 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2821 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2881 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2941 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3001 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3061 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3121 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     3181 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3241 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3301 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3361 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3421 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3481 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     3541 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3601 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3661 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3721 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3781 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3841 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3901 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     3961 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4021 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4081 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4141 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4201 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     4261 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4321 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4381 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4441 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4501 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     4561 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4621 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4681 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4741 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4801 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4861 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4921 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4981 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5041 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
