LOCUS       sequence-317299-        5045 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 5045)
  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 5045)
  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..5045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1774..1895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1811..1830)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1865..1884
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1902..2357)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2039..2060)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2251..2270
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2384..2488
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2490..2847
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2490..2681
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2513..2533)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2698..2833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2882..3676
                     /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             2882..3676
                     /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(2936..2955)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3546..3565
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3864..3883)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3908..3955
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4284..4872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4773..4792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcacc
      541 gccatgcaca gccagaagag ggtggtcgtg ctgggctccg gagtcatcgg gctctctagc
      601 gccctgattc tcgctagaaa aggctactcc gtgcatatcc tggccagaga cttgcctgag
      661 gatgtcagct cccagacctt cgcaagcccc tgggccggag ctaactggac accatttatg
      721 accctgacag acggacccag acaagccaag tgggaagagt ccaccttcaa gaaatgggtg
      781 gagctggttc ccacaggcca cgctatgtgg ctgaaaggga ccagaaggtt tgcccagaat
      841 gaggatggcc tcctgggaca ttggtacaag gacatcaccc ctaactatag gccacttcca
      901 agctctgaat gtccacctgg agccattgga gtgacatacg acaccctgag cgtccacgct
      961 cccaagtatt gccaatacct tgccagagag ctgcagaaac ttggggctac cttcgaaagg
     1021 agaacagtga ctagcctgga gcaagccttc gatggagctg acctcgtggt caacgccacc
     1081 gggctgggag ctaagtccat tgccggcatc gatgaccagg ctgccgagcc cattagggga
     1141 cagaccgtgc ttgtcaaatc tccctgtaag aggtgcacaa tggatagctc cgaccctgct
     1201 tctcccgctt atatcattcc tagaccagga ggagaagtga tctgtggcgg aacctacgga
     1261 gtcggcgatt gggacctgag cgtgaatccc gaaacagtcc aaaggattct caagcactgc
     1321 ctgagacttg accctaccat tagctccgat ggaactatcg agggcattga agtgctgaga
     1381 cataacgttg gcctgagacc cgccaggaga ggaggaccta gggtggaggc tgaaagaatc
     1441 gtcttgcccc tggaccggac caagagccct ctctctctgg gaagagggtc cgcacgggct
     1501 gcaaaagaga aggaagtgac ccttgtccac gcctacggct ttagtagcgc agggtatcag
     1561 caatcctggg gagccgctga ggacgtggct cagctggtcg atgaagcctt ccaaagatac
     1621 cacggcgcag ctagggagag ctgaggatcc accggatcta gataactgat cataatcagc
     1681 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1741 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1801 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1861 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     1921 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1981 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2041 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2101 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2161 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2221 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2281 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2341 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2401 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2461 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     2521 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2581 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2641 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2701 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2761 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2821 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     2881 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2941 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3001 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3061 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3121 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3181 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3241 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3301 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3361 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     3421 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3481 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3541 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3601 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3661 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3721 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3781 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3841 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3901 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     3961 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4021 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4081 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4141 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4201 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4261 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4321 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4381 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4441 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4501 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4561 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4621 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4681 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4741 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4801 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4861 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4921 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4981 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     5041 gttcc
//
