LOCUS       sequence-323598-        5248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DAAO with mitochondrial localization signal in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5248)
  TITLE     Mammalian expression plasmid for production of DAAO in the 
            mitochondrial matrix
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5248)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pC1-CMV-mito-DAAO
FEATURES             Location/Qualifiers
     source          1..5248
                     /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"
     CDS             597..671
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             693..764
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1902..1922
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     polyA_signal    2036..2157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2073..2092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2127..2146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2164..2619)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2301..2322)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2513..2532
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2646..2750
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2752..3109
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2752..2943
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2775..2795)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2960..3095
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3144..3938
                     /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             3144..3938
                     /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(3198..3217)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3808..3827
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4126..4145)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4170..4217
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4546..5134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5035..5054
                     /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 gctagcatgt
      601 ccgtcctgac gccgctgctg ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc
      661 cgcgcgccaa gatccattcg ttgggggatc tgtccgtcct gacgccgctg ctgctgcggg
      721 gcttgacagg ctcggcccgg cggctcccag tgccgcgcgc caagatccat tcgttggggg
      781 atccaccggt cgccacccac agccagaaga gggtggtcgt gctgggctcc ggagtcatcg
      841 ggctctctag cgccctgatt ctcgctagaa aaggctactc cgtgcatatc ctggccagag
      901 acttgcctga ggatgtcagc tcccagacct tcgcaagccc ctgggccgga gctaactgga
      961 caccgtttat gaccctgaca gacggaccca gacaagccaa gtgggaagag tccaccttca
     1021 agaaatgggt ggagctggtt cccacaggcc acgctatgtg gctgaaaggg accagaaggt
     1081 ttgcccagaa tgaggatggc ctcctgggac attggtacaa ggacatcacc cctaactata
     1141 ggccacttcc aagctctgaa tgtccacccg gagccattgg agtgacatac gacaccctga
     1201 gcgtccacgc tcccaagtat tgccaatacc ttgccagaga gctgcagaaa cttggggcta
     1261 ccttcgaaag gagaacagtg actagcctgg agcaagcctt cgatggagct gacctcgtgg
     1321 tcaacgccac cgggctggga gctaagtcca ttgccggcat cgatgaccag gctgccgagc
     1381 ccattagggg acagaccgtg cttgtcaaat ctccctgtaa gaggtgcaca atggatagct
     1441 ccgaccctgc ttctcccgct tatatcattc ctagaccagg aggagaagtg atctgtggcg
     1501 gaacctacgg agtcggcgat tgggacctga gcgtgaatcc cgaaacagtc caaaggattc
     1561 tcaagcactg cctgagactt gaccctacca ttagctccga tggaactatc gagggcattg
     1621 aagtgctgag acataacgtt ggcctgagac ccgccaggag aggaggacct agggtggagg
     1681 ctgaaagaat cgtcttgccc ctggaccgga ccaagagccc tctctctctg ggaagagggt
     1741 ccgcacgggc tgcaaaagag aaggaagtga cccttgtcca cgcctacggc tttagtagcg
     1801 cagggtatca gcaatcctgg ggagccgctg aggacgtggc tcagctggtc gatgaagcct
     1861 tccaaagata ccacggcgca gctagggaga gcgggggcgg gcatcatcat caccaccacc
     1921 attgatctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     1981 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     2041 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2101 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     2161 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     2221 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     2281 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     2341 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     2401 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2461 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2521 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2581 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     2641 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2701 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     2761 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2821 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2881 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2941 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3001 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3061 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     3121 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3181 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3241 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3301 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     3361 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3421 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3481 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3541 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3601 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3661 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3721 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3781 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3841 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3901 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3961 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4021 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4081 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     4141 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     4201 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4261 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4321 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4381 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4441 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4501 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4561 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4621 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4681 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4741 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4801 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4861 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4921 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4981 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5041 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5101 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5161 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5221 tgtggataac cgtattaccg ccatgcat
//
