LOCUS       sequence-430407-        6730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HyPerFLEX fused with UnaG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6730)
  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 6730)
  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-CMV-HyPerFLEX-UnaG-WPRE
FEATURES             Location/Qualifiers
     source          1..6730
                     /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        206..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     710..734
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          862..1240
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             2428..2844
                     /codon_start=1
                     /product="bilirubin-binding green fluorescent protein from 
                     Anguilla japonica (Kumagai et al., 2013)"
                     /label=UnaG
                     /translation="VEKFVGTWKIADSHNFGEYLKAIGAPKELSDGGDATTPTLYISQK
                     DGDKMTVKIENGPPTFLDTQVKFKLGEEFDEFPSDRRKGVKSVVNLVGEKLVYVQKWDG
                     KETTYVREIKDGKLVVTLTMGDVVAVRSYRRATE"
     misc_feature    2857..3445
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2910..2930)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3328..3339)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3477..3953
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3676..3695)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3993..4133
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3993..4122
                     /label=AAV2 ITR
     rep_origin      4208..4663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4295..4314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4505..4526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4680..4699)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4799..4821
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4859..4877)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4945..5049
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5050..5118,5119..5910)
                     /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(5268..5287)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6081..6669
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6570..6589
                     /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 gcgtctagtt attaatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      721 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      781 gggaccgatc cagcctccgc ggattcgaat cccggccggg aacggtgcat tggaacgcgg
      841 attccccgtg ccaagagtga cgtaagtacc gcctatagag tctataggcc cacaaaaaat
      901 gctttcttct tttaatatac ttttttgttt atcttatttc taatactttc cctaatctct
      961 ttctttcagg gcaataatga tacaatgtat catgcctctt tgcaccattc taaagaataa
     1021 cagtgataat ttctgggtta aggcaatagc aatatttctg catataaata tttctgcata
     1081 taaattgtaa ctgatgtaag aggtttcata ttgctaatag cagctacaat ccagctacca
     1141 ttctgctttt attttatggt tgggataagg ctggattatt ctgagtccaa gctaggccct
     1201 tttgctaatc atgttcatac ctcttatctt cctcccacag ctcctgggca acgtgctggt
     1261 ctgtgtgctg gcccatcact ttggcaaaga attgggattc gaacatcgat tgaattctga
     1321 gccaccatgc acctggctaa tgaggagcaa aacgaattgg aaggtgcttt taagctgggc
     1381 ttgatcttta ccgtcgcccc gtacttgctg cctaagttga tcgtcagcct gcgccgcacc
     1441 gccccgaaga tgcctttgat gttggaggag aattacaccc acacccttac cgagtcactg
     1501 aaacgcggcg acgttgacgc catcattgtg gcagagccgt ttcaagagcc gggtattgtg
     1561 actgaacctt tgtatgacga gccgtttttc gttatcgttc cgaagggcca tagcttcgag
     1621 gagttggatg ccgttagccc tcgcatgttg ggcgaagaac aagttctgct tctgaccgag
     1681 ggcaactgta tgcgtgacca ggtgcttagc atttttacca aggtcaaggt gcacatgaag
     1741 aaagcccttt ccggtgacag ctattgggtc tttgtgaaac gggtaggtgg cactggcggt
     1801 gagcatgttg tctttggcag tgaggacatc gagaacacta tggccaaaat ggacgacgga
     1861 caactggatg ggttggcctt tggcgcaatt cagctcgatg gtgacgggaa tatcctgcag
     1921 tacaatgctg ctgaaggaga catcacaggc agagatccca aacaggtgat tgggaagaac
     1981 ttcttcaagg atgttgcacc tggaacggat tctcccgagt tttacggcaa attcaaggaa
     2041 ggcgtagcgt cagggaatct gaacaccatg ttcgaatgga tgataccgac tcttaccgat
     2101 catagttgca gcgagcttgc cgccaaacag cgcattcagg gtctcaccaa cacccttcaa
     2161 ggctcaagca tcgataccat ccgccacatg gttgccagtg gcttggcaat cagcgtcttg
     2221 cctgccactg cccttaccga aaacgaccac atgctgttta gcattatccc gtttgaaggc
     2281 accccgccta gccgccgcgt tgtcctggcc tatcgccgca actttgtccg ccctaaggcc
     2341 ctttcagcca tgaaagccgc aattatgcaa agccaactgc atggtgttag cttcatctgc
     2401 gataccggtg tcgacccgcg gggatccgtc gagaaatttg ttggcacctg gaagatcgca
     2461 gacagccata attttggtga atacctgaaa gctatcggag ccccaaagga attaagcgat
     2521 ggtggggatg ccacgacgcc gacattgtac atctcccaga aggacggaga caaaatgaca
     2581 gtgaaaatag agaatggacc tcctacgttc cttgacactc aagtaaagtt caaattaggg
     2641 gaggagttcg acgaatttcc ttctgatcga agaaaaggcg taaaatctgt cgtgaacttg
     2701 gtgggagaga agctggtgta cgtacaaaag tgggacggca aggagacgac gtatgtccga
     2761 gagataaagg acggtaaact ggtcgtgaca cttacgatgg gagacgtcgt ggctgtgcgc
     2821 agctaccgga gggcgacgga atgaaagctt atcgataatc aacctctgga ttacaaaatt
     2881 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2941 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3001 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3061 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3121 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3181 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3241 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     3301 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3361 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3421 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt
     3481 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     3541 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     3601 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     3661 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     3721 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     3781 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     3841 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     3901 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     3961 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     4021 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     4081 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     4141 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     4201 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4261 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     4321 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4381 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     4441 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4501 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     4561 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     4621 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     4681 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4741 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4801 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4861 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4921 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4981 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5041 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5101 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5161 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5221 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5281 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5341 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5401 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5461 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5521 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5581 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5641 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5701 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5761 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5821 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5881 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5941 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6001 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6061 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6121 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6181 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6241 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6301 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6361 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6421 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6481 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6541 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6601 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6661 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6721 gctcacatgt
//
