LOCUS       sequence-481770-        6251 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expression of DAAO with a nuclear export signal and RFP as a 
            reporter in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6251)
  AUTHORS   Kalinichenko AL, Jappy D, Solius GM, Maltsev DI, Bogdanova YA, 
            Mukhametshina LF, Sokolov RA, Moshchenko AA, Shaydurov VA, Rozov AV,
            Podgorny OV, Belousov VV
  TITLE     Chemogenetic emulation of intraneuronal oxidative stress affects 
            synaptic plasticity.
  JOURNAL   Redox Biol. 2023 Apr;60:102604. doi: 10.1016/j.redox.2023.102604. 
            Epub 2023 Jan 10.
  PUBMED    36640726
REFERENCE   2  (bases 1 to 6251)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Redox 
            Biol."; date: "2023-04"; volume: "60:102604. doi"; pages: " 
            10.1016/j.redox.2023.102604. Epub 2023 Jan 10"
COMMENT     Sequence Label: pAAV_CMV-RFP-p2A-DAAO-NES
FEATURES             Location/Qualifiers
     source          1..6251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        145..448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        449..652
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     603..623
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      679..688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             685..1395
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             1405..1461
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2557..2583
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     misc_feature    2597..3185
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2650..2670)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3068..3079)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3354..3435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3391..3410)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3445..3464
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3514..3654
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3514..3643
                     /label=AAV2 ITR
     rep_origin      3729..4184
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3816..3835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4026..4047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4201..4220)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4320..4342
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4380..4398)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4466..4570
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4571..4639,4640..5431)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4789..4808)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5602..6190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6091..6110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcgttac ataacttacg gtaaatggcc cgcctggctg
      181 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      241 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      301 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      361 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      421 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      481 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      541 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      601 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      661 gaaccgtcag atccgctagc caccatggtg tctaagggcg aagagctgat taaggagaac
      721 atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa gtgcacatcc
      781 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc
      841 ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg cagcagaacc
      901 ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc tgagggcttc
      961 acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc
     1021 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcccatcc
     1081 aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccaacaccga gatgctgtac
     1141 cccgctgacg gcggcctgga aggcagaagc gacatggccc tgaagctcgt gggcgggggc
     1201 cacctgatct gcaacttcaa gaccacatac agatccaaga aacccgctaa gaacctcaag
     1261 atgcccggcg tctactatgt ggaccacaga ctggaaagaa tcaaggaggc cgacaaagag
     1321 acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc tagcaaactg
     1381 gggcacaaac ttaatggatc cggagccacg aacttctctc tgttaaagca agcaggagac
     1441 gtggaagaaa accccggtcc tcacagccag aagagggtgg tcgtgctggg ctccggagtc
     1501 atcgggctct ctagcgccct gattctcgct agaaaaggct actccgtgca tatcctggcc
     1561 agagacttgc ctgaggatgt cagctcccag accttcgcaa gcccctgggc cggagctaac
     1621 tggacaccat ttatgaccct gacagacgga cccagacaag ccaagtggga agagtccacc
     1681 ttcaagaaat gggtggagct ggttcccaca ggccacgcta tgtggctgaa agggaccaga
     1741 aggtttgccc agaatgagga tggcctcctg ggacattggt acaaggacat cacccctaac
     1801 tataggccac ttccaagctc tgaatgtcca cctggagcca ttggagtgac atacgacacc
     1861 ctgagcgtcc acgctcccaa gtattgccaa taccttgcca gagagctgca gaaacttggg
     1921 gctaccttcg aaaggagaac agtgactagc ctggagcaag ccttcgatgg agctgacctc
     1981 gtggtcaacg ccaccgggct gggagctaag tccattgccg gcatcgatga ccaggctgcc
     2041 gagcccatta ggggacagac cgtgcttgtc aaatctccct gtaagaggtg cacaatggat
     2101 agctccgacc ctgcttctcc cgcttatatc attcctagac caggaggaga agtgatctgt
     2161 ggcggaacct acggagtcgg cgattgggac ctgagcgtga atcccgaaac agtccaaagg
     2221 attctcaagc actgcctgag acttgaccct accattagct ccgatggaac tatcgagggc
     2281 attgaagtgc tgagacataa cgttggcctg agacccgcca ggagaggagg acctagggtg
     2341 gaggctgaaa gaatcgtctt gcccctggac cggaccaaga gccctctctc tctgggaaga
     2401 gggtccgcac gggctgcaaa agagaaggaa gtgacccttg tccacgccta cggctttagt
     2461 agcgcagggt atcagcaatc ctggggagcc gctgaggacg tggctcagct ggtcgatgaa
     2521 gccttccaaa gataccacgg cgcagctagg gagagcctgc cccccctgga gcggctgacc
     2581 ctgtaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2641 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2701 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2761 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2821 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2881 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2941 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     3001 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     3061 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3121 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3181 cccgcatcga taccgagcgc tgctcgagag atctgtgata gcggccatca agctggccgc
     3241 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3301 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3361 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3421 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcagcttatc
     3481 gataccgcat gcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     3541 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3601 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     3661 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     3721 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3781 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     3841 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3901 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     3961 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4021 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4081 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4141 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4201 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4261 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4321 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4381 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4441 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4501 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4561 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4621 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4681 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4741 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4801 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4861 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4921 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4981 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5041 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5101 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5161 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5221 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5281 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5341 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5401 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5461 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5521 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5581 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5641 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5701 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5761 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5821 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5881 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5941 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6001 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6061 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6121 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6181 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6241 tgctcacatg t
//
