LOCUS       sequence-430367-        5688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HyPerFLEX targeted to the cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5688)
  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 5688)
  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-CAG-HyPerFLEX-NES-WPRE
FEATURES             Location/Qualifiers
     source          1..5688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..65
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   224..353
                     /label=AAV2 ITR
     primer_bind     1228..1250
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1312..1331
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2462..2488
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     misc_feature    2504..3092
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2557..2577)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2975..2986)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3135..3256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3146..3165)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3200..3219
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3635..4223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3715..3734)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4394..5185,5186..5254))
                     /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     5017..5036
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5255..5359)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5385..5688,1..152))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5522..5543)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
       61 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
      121 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
      181 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg ctcgctcgct
      241 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
      301 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt
      361 aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga tcgtaccatt
      421 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      481 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      541 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      601 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      661 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      721 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
      781 gggggggngc gcgccnrggs ggggnnsggg gggggggggg gggggggggg gggggggrgg
      841 sggrgaggkg sggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      901 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
      961 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
     1021 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
     1081 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
     1141 ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg gggnacggct
     1201 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     1261 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     1321 tggttattgt gctgtctcat cattttggca aagaattgga tcccatcgat tcgaattcgc
     1381 caccatgcac ctggctaatg aggagcaaaa cgaattggaa ggtgctttta agctgggctt
     1441 gatctttacc gtcgccccgt acttgctgcc taagttgatc gtcagcctgc gccgcaccgc
     1501 cccgaagatg cctttgatgt tggaggagaa ttacacccac acccttaccg agtcactgaa
     1561 acgcggcgac gttgacgcca tcattgtggc agagccgttt caagagccgg gtattgtgac
     1621 tgaacctttg tatgacgagc cgtttttcgt tatcgttccg aagggccata gcttcgagga
     1681 gttggatgcc gttagccctc gcatgttggg cgaagaacaa gttctgcttc tgaccgaggg
     1741 caactgtatg cgtgaccagg tgcttagcat ttttaccaag gtcaaggtgc acatgaagaa
     1801 agccctttcc ggtgacagct attgggtctt tgtgaaacgg gtaggtggca ctggcggtga
     1861 gcatgttgtc tttggcagtg aggacatcga gaacactatg gccaaaatgg acgacggaca
     1921 actggatggg ttggcctttg gcgcaattca gctcgatggt gacgggaata tcctgcagta
     1981 caatgctgct gaaggagaca tcacaggcag agatcccaaa caggtgattg ggaagaactt
     2041 cttcaaggat gttgcacctg gaacggattc tcccgagttt tacggcaaat tcaaggaagg
     2101 cgtagcgtca gggaatctga acaccatgtt cgaatggatg ataccgactc ttaccgatca
     2161 tagttgcagc gagcttgccg ccaaacagcg cattcagggt ctcaccaaca cccttcaagg
     2221 ctcaagcatc gataccatcc gccacatggt tgccagtggc ttggcaatca gcgtcttgcc
     2281 tgccactgcc cttaccgaaa acgaccacat gctgtttagc attatcccgt ttgaaggcac
     2341 cccgcctagc cgccgcgttg tcctggccta tcgccgcaac tttgtccgcc ctaaggccct
     2401 ttcagccatg aaagccgcaa ttatgcaaag ccaactgcat ggtgttagct tcatctgcga
     2461 tctgcccccc ctggagcggc tgaccctgta aaagcttatc gataatcaac ctctggatta
     2521 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2581 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2641 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2701 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2761 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2821 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2881 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     2941 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3001 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3061 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct
     3121 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3181 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3241 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3301 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     3361 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     3421 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgactcgc
     3481 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3541 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3601 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3661 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3721 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3781 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3841 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3901 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3961 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4021 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4081 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4141 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4201 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4261 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4321 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4381 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4441 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4501 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4561 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4621 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4681 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4741 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4801 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4861 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4921 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4981 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5041 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5101 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5161 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5221 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5281 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5341 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     5401 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     5461 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5521 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5581 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5641 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccg
//
