LOCUS       sequence-483824-        7012 bp    DNA     circular SYN 19-AUG-2025
DEFINITION  Mammalian expression of mutated DAAO(R285A) with a nuclear export 
            signal and RFP as a reporter in excitatory neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7012)
  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 7012)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 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_CaMKII-RFP-p2A-DAAO(R285A)-NES
FEATURES             Location/Qualifiers
     source          1..7012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..126
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        145..1433
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1438..1447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1444..2154
                     /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             2164..2220
                     /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             3316..3342
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     misc_feature    3358..3946
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3411..3431)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3829..3840)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4115..4196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4152..4171)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4206..4225
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4275..4415
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4275..4404
                     /label=AAV2 ITR
     rep_origin      4490..4945
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4577..4596)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4787..4808
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4962..4981)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5081..5103
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5141..5159)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5227..5331
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5332..5400,5401..6192)
                     /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(5550..5569)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6363..6951
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6852..6871
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      121 gttcctgcgg ccgcacgcgt ttaacattat ggccttaggt cacttcatct ccatggggtt
      181 cttcttctga ttttctagaa aatgagatgg gggtgcagag agcttcctca gtgacctgcc
      241 cagggtcaca tcagaaatgt cagagctaga acttgaactc agattactaa tcttaaattc
      301 catgccttgg gggcatgcaa gtacgatata cagaaggagt gaactcatta gggcagatga
      361 ccaatgagtt taggaaagaa gagtccaggg cagggtacat ctacaccacc cgcccagccc
      421 tgggtgagtc cagccacgtt cacctcatta tagttgcctc tctccagtcc taccttgacg
      481 ggaagcacaa gcagaaactg ggacaggagc cccaggagac caaatcttca tggtccctct
      541 gggaggatgg gtggggagag ctgtggcaga ggcctcagga ggggccctgc tgctcagtgg
      601 tgacagatag gggtgagaaa gcagacagag tcattccgtc agcattctgg gtctgtttgg
      661 tacttcttct cacgctaagg tggcggtgtg atatgcacaa tggctaaaaa gcagggagag
      721 ctggaaagaa acaaggacag agacagaggc caagtcaacc agaccaattc ccagaggaag
      781 caaagaaacc attacagaga ctacaagggg gaagggaagg agagatgaat tagcttcccc
      841 tgtaaacctt agaacccagc tgttgccagg gcaacggggc aatacctgtc tcttcagagg
      901 agatgaagtt gccagggtaa ctacatcctg tctttctcaa ggaccatccc agaatgtggc
      961 acccactagc cgttaccata gcaactgcct ctttgcccca cttaatccca tcccgtctgt
     1021 taaaagggcc ctatagttgg aggtggggga ggtaggaaga gcgatgatca cttgtggact
     1081 aagtttgttc gcatcccctt ctccaacccc ctcagtacat caccctgggg gaacagggtc
     1141 cacttgctcc tgggcccaca cagtcctgca gtattgtgta tataaggcca gggcaaagag
     1201 gagcaggttt taaagtgaaa ggcaggcagg tgttggggag gcagttaccg gggcaacggg
     1261 aacagggcgt ttcggaggtg gttgccatgg ggacctggat gctgacgaag gctcgcgagg
     1321 ctgtgagcag ccacagtgcc ctgctcagaa gccccaagct cgtcagtcaa gccggttctc
     1381 cgtttgcact caggagcacg ggcaggcgag tggcccctag ttctgggggc agcgctagcc
     1441 accatggtgt ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg
     1501 gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac
     1561 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
     1621 gacatcctgg ctaccagctt catgtacggc agcagaacct tcatcaacca cacccagggc
     1681 atccccgact tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca
     1741 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
     1801 atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag
     1861 aaaacactcg gctgggaggc caacaccgag atgctgtacc ccgctgacgg cggcctggaa
     1921 ggcagaagcg acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttcaag
     1981 accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg
     2041 gaccacagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag
     2101 gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacaaact taatggatcc
     2161 ggagccacga acttctctct gttaaagcaa gcaggagacg tggaagaaaa ccccggtcct
     2221 cacagccaga agagggtggt cgtgctgggc tccggagtca tcgggctctc tagcgccctg
     2281 attctcgcta gaaaaggcta ctccgtgcat atcctggcca gagacttgcc tgaggatgtc
     2341 agctcccaga ccttcgcaag cccctgggcc ggagctaact ggacaccatt tatgaccctg
     2401 acagacggac ccagacaagc caagtgggaa gagtccacct tcaagaaatg ggtggagctg
     2461 gttcccacag gccacgctat gtggctgaaa gggaccagaa ggtttgccca gaatgaggat
     2521 ggcctcctgg gacattggta caaggacatc acccctaact ataggccact tccaagctct
     2581 gaatgtccac ctggagccat tggagtgaca tacgacaccc tgagcgtcca cgctcccaag
     2641 tattgccaat accttgccag agagctgcag aaacttgggg ctaccttcga aaggagaaca
     2701 gtgactagcc tggagcaagc cttcgatgga gctgacctcg tggtcaacgc caccgggctg
     2761 ggagctaagt ccattgccgg catcgatgac caggctgccg agcccattag gggacagacc
     2821 gtgcttgtca aatctccctg taagaggtgc acaatggata gctccgaccc tgcttctccc
     2881 gcttatatca ttcctagacc aggaggagaa gtgatctgtg gcggaaccta cggagtcggc
     2941 gattgggacc tgagcgtgaa tcccgaaaca gtccaaagga ttctcaagca ctgcctgaga
     3001 cttgacccta ccattagctc cgatggaact atcgagggca ttgaagtgct gagacataac
     3061 gttggcctgg cacccgccag gagaggagga cctagggtgg aggctgaaag aatcgtcttg
     3121 cccctggacc ggaccaagag ccctctctct ctgggaagag ggtccgcacg ggctgcaaaa
     3181 gagaaggaag tgacccttgt ccacgcctac ggctttagta gcgcagggta tcagcaatcc
     3241 tggggagccg ctgaggacgt ggctcagctg gtcgatgaag ccttccaaag ataccacggc
     3301 gcagctaggg agagcctgcc ccccctggag cggctgaccc tgtaaaagct tatcgataat
     3361 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3421 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3481 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3541 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3601 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3661 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3721 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     3781 tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3841 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3901 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg
     3961 ctgctcgaga gatctgtgat agcggccatc aagctggccg cgactctaga tcataatcag
     4021 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     4081 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     4141 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4201 tagttgtggt ttgtccaaac tcatcaatgt atcagcttat cgataccgca tgcacgtgcg
     4261 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     4321 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     4381 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     4441 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     4501 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4561 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4621 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4681 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     4741 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4801 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     4861 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4921 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     4981 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5041 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5101 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     5161 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5221 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5281 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5341 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5401 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5461 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5521 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5581 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5641 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5701 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5761 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5821 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5881 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5941 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6001 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6061 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6121 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6181 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6241 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6301 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6361 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6421 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6481 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6541 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6601 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6661 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6721 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6781 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6841 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6901 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6961 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
