LOCUS       sequence-430410-        7026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of DAAO targeted to the cytoplasm and mTagBFP2, separated
            by P2A self-cleaving peptide, in mammalian neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7026)
  AUTHORS   Potekhina ES, Bass DI, Ezerina D, Fleckenstein DD, Chebotarev AS, 
            Sysoeva VA, Maltsev DI, Pak VV, Moshchenko AA, Sokolov AI, 
            Myasnyanko IN, Baranov MS, Fedotov AB, Zheltikov AM, Lanin AA, 
            Messens J, Belousov VV
  TITLE     A color-tailored fluorogenic sensor for hydrogen peroxide.
  JOURNAL   Nat Chem Biol. 2025 Oct 16. doi: 10.1038/s41589-025-02036-6.
  PUBMED    41102408
REFERENCE   2  (bases 1 to 7026)
  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: "Nat Chem 
            Biol. 2025 Oct 16. doi: 10.1038/s41589-025-02036-6."
COMMENT     Sequence Label: pAAV-CaMKII-mTagBFP2-P2A-DAAO-NES
FEATURES             Location/Qualifiers
     source          1..7026
                     /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      1454..1463
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1460..2170
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             2180..2236
                     /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             3332..3358
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     misc_feature    3372..3960
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3425..3445)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3843..3854)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4129..4210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4166..4185)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4220..4239
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4289..4429
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4289..4418
                     /label=AAV2 ITR
     rep_origin      4504..4959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4591..4610)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4801..4822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4976..4995)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5095..5117
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5155..5173)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5241..5345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5346..5414,5415..6206)
                     /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(5564..5583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6377..6965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6866..6885
                     /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 agcgctaccg gtcgccacca tggtgtctaa gggcgaagag ctgattaagg agaacatgca
     1501 catgaagctg tacatggagg gcaccgtgga caaccatcac ttcaagtgca catccgaggg
     1561 cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc
     1621 tctccccttc gccttcgaca tcctggctac tagcttcctc tacggcagca agaccttcat
     1681 caaccacacc cagggcatcc ccgacttctt caagcagtcc ttccctgagg gcttcacatg
     1741 ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct
     1801 ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttca catccaacgg
     1861 ccctgtgatg cagaagaaaa cactcggctg ggaggccttc accgagacgc tgtaccccgc
     1921 tgacggcggc ctggaaggca gaaacgacat ggccctgaag ctcgtgggcg ggagccatct
     1981 gatcgcaaac gccaagacca catatagatc caagaaaccc gctaagaacc tcaagatgcc
     2041 tggcgtctac tatgtggact acagactgga aagaatcaag gaggccaaca acgagaccta
     2101 cgtcgagcag cacgaggtgg cagtggccag atactgcgac ctccctagca aactggggca
     2161 caagcttaat ggatccggag ccacgaactt ctctctgtta aagcaagcag gagacgtgga
     2221 agaaaacccc ggtcctcaca gccagaagag ggtggtcgtg ctgggctccg gagtcatcgg
     2281 gctctctagc gccctgattc tcgctagaaa aggctactcc gtgcatatcc tggccagaga
     2341 cttgcctgag gatgtcagct cccagacctt cgcaagcccc tgggccggag ctaactggac
     2401 accatttatg accctgacag acggacccag acaagccaag tgggaagagt ccaccttcaa
     2461 gaaatgggtg gagctggttc ccacaggcca cgctatgtgg ctgaaaggga ccagaaggtt
     2521 tgcccagaat gaggatggcc tcctgggaca ttggtacaag gacatcaccc ctaactatag
     2581 gccacttcca agctctgaat gtccacctgg agccattgga gtgacatacg acaccctgag
     2641 cgtccacgct cccaagtatt gccaatacct tgccagagag ctgcagaaac ttggggctac
     2701 cttcgaaagg agaacagtga ctagcctgga gcaagccttc gatggagctg acctcgtggt
     2761 caacgccacc gggctgggag ctaagtccat tgccggcatc gatgaccagg ctgccgagcc
     2821 cattagggga cagaccgtgc ttgtcaaatc tccctgtaag aggtgcacaa tggatagctc
     2881 cgaccctgct tctcccgctt atatcattcc tagaccagga ggagaagtga tctgtggcgg
     2941 aacctacgga gtcggcgatt gggacctgag cgtgaatccc gaaacagtcc aaaggattct
     3001 caagcactgc ctgagacttg accctaccat tagctccgat ggaactatcg agggcattga
     3061 agtgctgaga cataacgttg gcctgagacc cgccaggaga ggaggaccta gggtggaggc
     3121 tgaaagaatc gtcttgcccc tggaccggac caagagccct ctctctctgg gaagagggtc
     3181 cgcacgggct gcaaaagaga aggaagtgac ccttgtccac gcctacggct ttagtagcgc
     3241 agggtatcag caatcctggg gagccgctga ggacgtggct cagctggtcg atgaagcctt
     3301 ccaaagatac cacggcgcag ctagggagag cctgcccccc ctggagcggc tgaccctgta
     3361 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3421 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3481 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3541 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3601 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3661 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3721 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     3781 cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3841 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3901 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3961 atcgataccg agcgctgctc gagagatctg tgatagcggc catcaagctg gccgcgactc
     4021 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4081 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4141 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4201 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatcagc ttatcgatac
     4261 cgcatgcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4321 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4381 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4441 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4501 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4561 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4621 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4681 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4741 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4801 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     4861 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     4921 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4981 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5041 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5101 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5161 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5221 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5281 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5341 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5401 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5461 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5521 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5581 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5641 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5701 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5761 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5821 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5881 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5941 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6001 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6061 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6121 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6181 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6241 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6301 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6361 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6421 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6481 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6541 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6601 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6661 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6721 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6781 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6841 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6901 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6961 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7021 acatgt
//
