LOCUS       sequence-430400-        6632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HyPerFLEX fused with UnaG in mammalian neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6632)
  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 6632)
  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: AAV-CaMKII-HyPerFLEX-UnaG-WPRE
FEATURES             Location/Qualifiers
     source          1..6632
                     /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"
     CDS             2549..2965
                     /codon_start=1
                     /product="bilirubin-binding green fluorescent protein from 
                     Anguilla japonica (Kumagai et al., 2013)"
                     /label=UnaG
                     /translation="VEKFVGTWKIADSHNFGEYLKAIGAPKELSDGGDATTPTLYISQK
                     DGDKMTVKIENGPPTFLDTQVKFKLGEEFDEFPSDRRKGVKSVVNLVGEKLVYVQKWDG
                     KETTYVREIKDGKLVVTLTMGDVVAVRSYRRATE"
     misc_feature    2978..3566
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3031..3051)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3449..3460)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3735..3816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3772..3791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3826..3845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3895..4035
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3895..4024
                     /label=AAV2 ITR
     rep_origin      4110..4565
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4197..4216)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4407..4428
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4582..4601)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4701..4723
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4761..4779)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4847..4951
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4952..5020,5021..5812)
                     /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(5170..5189)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5983..6571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6472..6491
                     /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 cacctggcta atgaggagca aaacgaattg gaaggtgctt ttaagctggg
     1501 cttgatcttt accgtcgccc cgtacttgct gcctaagttg atcgtcagcc tgcgccgcac
     1561 cgccccgaag atgcctttga tgttggagga gaattacacc cacaccctta ccgagtcact
     1621 gaaacgcggc gacgttgacg ccatcattgt ggcagagccg tttcaagagc cgggtattgt
     1681 gactgaacct ttgtatgacg agccgttttt cgttatcgtt ccgaagggcc atagcttcga
     1741 ggagttggat gccgttagcc ctcgcatgtt gggcgaagaa caagttctgc ttctgaccga
     1801 gggcaactgt atgcgtgacc aggtgcttag catttttacc aaggtcaagg tgcacatgaa
     1861 gaaagccctt tccggtgaca gctattgggt ctttgtgaaa cgggtaggtg gcactggcgg
     1921 tgagcatgtt gtctttggca gtgaggacat cgagaacact atggccaaaa tggacgacgg
     1981 acaactggat gggttggcct ttggcgcaat tcagctcgat ggtgacggga atatcctgca
     2041 gtacaatgct gctgaaggag acatcacagg cagagatccc aaacaggtga ttgggaagaa
     2101 cttcttcaag gatgttgcac ctggaacgga ttctcccgag ttttacggca aattcaagga
     2161 aggcgtagcg tcagggaatc tgaacaccat gttcgaatgg atgataccga ctcttaccga
     2221 tcatagttgc agcgagcttg ccgccaaaca gcgcattcag ggtctcacca acacccttca
     2281 aggctcaagc atcgatacca tccgccacat ggttgccagt ggcttggcaa tcagcgtctt
     2341 gcctgccact gcccttaccg aaaacgacca catgctgttt agcattatcc cgtttgaagg
     2401 caccccgcct agccgccgcg ttgtcctggc ctatcgccgc aactttgtcc gccctaaggc
     2461 cctttcagcc atgaaagccg caattatgca aagccaactg catggtgtta gcttcatctg
     2521 cgataccggt gtcgacccgc ggggatccgt cgagaaattt gttggcacct ggaagatcgc
     2581 agacagccat aattttggtg aatacctgaa agctatcgga gccccaaagg aattaagcga
     2641 tggtggggat gccacgacgc cgacattgta catctcccag aaggacggag acaaaatgac
     2701 agtgaaaata gagaatggac ctcctacgtt ccttgacact caagtaaagt tcaaattagg
     2761 ggaggagttc gacgaatttc cttctgatcg aagaaaaggc gtaaaatctg tcgtgaactt
     2821 ggtgggagag aagctggtgt acgtacaaaa gtgggacggc aaggagacga cgtatgtccg
     2881 agagataaag gacggtaaac tggtcgtgac acttacgatg ggagacgtcg tggctgtgcg
     2941 cagctaccgg agggcgacgg aatgaaagct tatcgataat caacctctgg attacaaaat
     3001 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3061 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3121 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3181 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3241 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3301 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3361 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     3421 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3481 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3541 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctgtgat
     3601 agcggccatc aagctggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     3661 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3721 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3781 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3841 tcatcaatgt atcagcttat cgataccgca tgcacgtgcg gaccgagcgg ccgcaggaac
     3901 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     3961 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     4021 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4081 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     4141 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     4201 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4261 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4321 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4381 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4441 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     4501 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4561 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4621 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4681 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4741 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4801 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4861 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4921 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4981 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5041 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5101 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5161 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5221 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5281 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5341 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5401 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5461 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5521 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5581 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5641 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5701 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5761 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5821 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5881 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5941 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6001 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6061 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6121 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6181 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6241 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6301 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6361 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6421 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6481 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6541 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6601 cctggccttt tgctggcctt ttgctcacat gt
//
