LOCUS       sequence-430397-        5724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HyPerFLEX targeted to the nucleus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5724)
  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 5724)
  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-NLSx3-WPRE
FEATURES             Location/Qualifiers
     source          1..5724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..55
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   214..343
                     /label=AAV2 ITR
     promoter        656..933
                     /label=chicken beta-actin promoter
     primer_bind     1212..1234
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1296..1315
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2446..2466
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2470..2490
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2494..2514
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2530..3118
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2583..2603)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3001..3012)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3161..3282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3172..3191)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3226..3245
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3661..4249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3741..3760)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4420..5211,5212..5280))
                     /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     5043..5062
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5281..5385)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5411..5724,1..142))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5548..5569)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
       61 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
      121 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
      181 cgatcggtgc gggcctcttc gctattacgc cagctgcgcg ctcgctcgct cactgaggcc
      241 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
      301 gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac
      361 ccgccatgct acttatctac gtagccatgc tctaggaaga tcgtaccatt gacgtcaata
      421 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      481 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      541 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      601 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
      661 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
      721 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc
      781 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
      841 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
      901 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
      961 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
     1021 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
     1081 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct
     1141 ttgtgcgggg ggagcggctc ggggctgtcc gcggggggac ggctgccttc gggggggacg
     1201 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
     1261 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
     1321 tcatcatttt ggcaaagaat tggatcccat cgattcgaat tcgccaccat gcacctggct
     1381 aatgaggagc aaaacgaatt ggaaggtgct tttaagctgg gcttgatctt taccgtcgcc
     1441 ccgtacttgc tgcctaagtt gatcgtcagc ctgcgccgca ccgccccgaa gatgcctttg
     1501 atgttggagg agaattacac ccacaccctt accgagtcac tgaaacgcgg cgacgttgac
     1561 gccatcattg tggcagagcc gtttcaagag ccgggtattg tgactgaacc tttgtatgac
     1621 gagccgtttt tcgttatcgt tccgaagggc catagcttcg aggagttgga tgccgttagc
     1681 cctcgcatgt tgggcgaaga acaagttctg cttctgaccg agggcaactg tatgcgtgac
     1741 caggtgctta gcatttttac caaggtcaag gtgcacatga agaaagccct ttccggtgac
     1801 agctattggg tctttgtgaa acgggtaggt ggcactggcg gtgagcatgt tgtctttggc
     1861 agtgaggaca tcgagaacac tatggccaaa atggacgacg gacaactgga tgggttggcc
     1921 tttggcgcaa ttcagctcga tggtgacggg aatatcctgc agtacaatgc tgctgaagga
     1981 gacatcacag gcagagatcc caaacaggtg attgggaaga acttcttcaa ggatgttgca
     2041 cctggaacgg attctcccga gttttacggc aaattcaagg aaggcgtagc gtcagggaat
     2101 ctgaacacca tgttcgaatg gatgataccg actcttaccg atcatagttg cagcgagctt
     2161 gccgccaaac agcgcattca gggtctcacc aacacccttc aaggctcaag catcgatacc
     2221 atccgccaca tggttgccag tggcttggca atcagcgtct tgcctgccac tgcccttacc
     2281 gaaaacgacc acatgctgtt tagcattatc ccgtttgaag gcaccccgcc tagccgccgc
     2341 gttgtcctgg cctatcgccg caactttgtc cgccctaagg ccctttcagc catgaaagcc
     2401 gcaattatgc aaagccaact gcatggtgtt agcttcatct gcgatccaaa aaagaagaga
     2461 aaggtagatc caaaaaagaa gagaaaggta gatccaaaaa agaagagaaa ggtataaaag
     2521 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2581 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2641 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2701 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2761 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2821 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2881 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     2941 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3001 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3061 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3121 cgataccgtc gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt
     3181 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3241 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3301 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc
     3361 tacaaatgtg gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca
     3421 tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct
     3481 gcgcgctcgc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3541 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3601 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3661 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3721 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3781 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3841 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3901 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3961 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4021 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4081 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4141 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4201 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4261 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4321 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4381 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4441 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4501 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4561 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4621 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4681 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4741 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4801 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4861 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4921 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4981 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5041 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5101 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5161 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5221 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5281 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5341 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5401 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     5461 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     5521 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     5581 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     5641 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     5701 aaagggagcc cccgatttag agct
//
