LOCUS       sequence-481776-        7014 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Mammalian expression of DAAO 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 7014)
  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 7014)
  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_CaMKII-RFP-p2A-DAAO-NES
FEATURES             Location/Qualifiers
     source          1..7014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1442..1451
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1448..2158
                     /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             2168..2224
                     /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             3320..3346
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     misc_feature    3360..3948
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3413..3433)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3831..3842)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4117..4198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4154..4173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4208..4227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4277..4417
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4277..4406
                     /label=AAV2 ITR
     rep_origin      4492..4947
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4579..4598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4789..4810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4964..4983)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5083..5105
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5143..5161)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5229..5333
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5334..5402,5403..6194)
                     /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(5552..5571)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6365..6953
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6854..6873
                     /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 gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagcgct
     1441 agccaccatg gtgtctaagg gcgaagagct gattaaggag aacatgcaca tgaagctgta
     1501 catggagggc accgtgaaca accaccactt caagtgcaca tccgagggcg aaggcaagcc
     1561 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc
     1621 cttcgacatc ctggctacca gcttcatgta cggcagcaga accttcatca accacaccca
     1681 gggcatcccc gacttcttta agcagtcctt ccctgagggc ttcacatggg agagagtcac
     1741 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg
     1801 cctcatctac aacgtcaaga tcagaggggt gaacttccca tccaacggcc ctgtgatgca
     1861 gaagaaaaca ctcggctggg aggccaacac cgagatgctg taccccgctg acggcggcct
     1921 ggaaggcaga agcgacatgg ccctgaagct cgtgggcggg ggccacctga tctgcaactt
     1981 caagaccaca tacagatcca agaaacccgc taagaacctc aagatgcccg gcgtctacta
     2041 tgtggaccac agactggaaa gaatcaagga ggccgacaaa gagacctacg tcgagcagca
     2101 cgaggtggct gtggccagat actgcgacct ccctagcaaa ctggggcaca aacttaatgg
     2161 atccggagcc acgaacttct ctctgttaaa gcaagcagga gacgtggaag aaaaccccgg
     2221 tcctcacagc cagaagaggg tggtcgtgct gggctccgga gtcatcgggc tctctagcgc
     2281 cctgattctc gctagaaaag gctactccgt gcatatcctg gccagagact tgcctgagga
     2341 tgtcagctcc cagaccttcg caagcccctg ggccggagct aactggacac catttatgac
     2401 cctgacagac ggacccagac aagccaagtg ggaagagtcc accttcaaga aatgggtgga
     2461 gctggttccc acaggccacg ctatgtggct gaaagggacc agaaggtttg cccagaatga
     2521 ggatggcctc ctgggacatt ggtacaagga catcacccct aactataggc cacttccaag
     2581 ctctgaatgt ccacctggag ccattggagt gacatacgac accctgagcg tccacgctcc
     2641 caagtattgc caataccttg ccagagagct gcagaaactt ggggctacct tcgaaaggag
     2701 aacagtgact agcctggagc aagccttcga tggagctgac ctcgtggtca acgccaccgg
     2761 gctgggagct aagtccattg ccggcatcga tgaccaggct gccgagccca ttaggggaca
     2821 gaccgtgctt gtcaaatctc cctgtaagag gtgcacaatg gatagctccg accctgcttc
     2881 tcccgcttat atcattccta gaccaggagg agaagtgatc tgtggcggaa cctacggagt
     2941 cggcgattgg gacctgagcg tgaatcccga aacagtccaa aggattctca agcactgcct
     3001 gagacttgac cctaccatta gctccgatgg aactatcgag ggcattgaag tgctgagaca
     3061 taacgttggc ctgagacccg ccaggagagg aggacctagg gtggaggctg aaagaatcgt
     3121 cttgcccctg gaccggacca agagccctct ctctctggga agagggtccg cacgggctgc
     3181 aaaagagaag gaagtgaccc ttgtccacgc ctacggcttt agtagcgcag ggtatcagca
     3241 atcctgggga gccgctgagg acgtggctca gctggtcgat gaagccttcc aaagatacca
     3301 cggcgcagct agggagagcc tgccccccct ggagcggctg accctgtaag cttatcgata
     3361 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3421 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3481 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3541 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3601 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3661 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3721 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3781 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3841 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3901 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     3961 cgctgctcga gagatctgtg atagcggcca tcaagctggc cgcgactcta gatcataatc
     4021 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4081 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4141 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4201 tctagttgtg gtttgtccaa actcatcaat gtatcagctt atcgataccg catgcacgtg
     4261 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4321 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     4381 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     4441 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     4501 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4561 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4621 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4681 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4741 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4801 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     4861 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4921 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     4981 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5041 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5101 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     5161 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5221 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5281 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5341 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5401 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5461 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5521 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5581 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5641 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5701 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5761 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5821 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5881 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5941 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6001 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6061 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6121 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6181 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6241 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6301 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6361 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6421 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6481 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6541 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6601 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6661 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6721 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6781 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6841 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6901 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6961 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
