LOCUS       sequence-430422-        5237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression HyPerFLEX targeted to actin filaments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5237)
  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 5237)
  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: pSC2+-HyPerFLEX-Lifeact
FEATURES             Location/Qualifiers
     source          1..5237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1198..1248
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     polyA_signal    1289..1423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1313..1332)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1367..1386
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1536..1556)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1536..1554)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1575..1591)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1575..1591)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1588..1610)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1599..1615
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1623..1629,1630..1647,1648..1653))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1668..1689
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1806..1823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1977..2565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2057..2076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2736..3527,3528..3596))
                     /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     3359..3378
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3597..3701)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3727..4182)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3864..3885)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4076..4095
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4253..5237
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattcg ccaccatgca cctggccaat
      121 gaggagcaaa acgaattgga aggtgctttt aagctgggct tgatctttac cgtcgccccg
      181 tacttgctgc ctaagttgat cgtcagcctg cgccgcaccg ccccgaagat gcctttgatg
      241 ttggaggaga attacaccca cacccttacc gagtcactga aacgcggcga cgttgacgcc
      301 atcattgtgg cagagccgtt tcaagagccg ggtattgtga ctgaaccttt gtatgacgag
      361 ccgtttttcg ttatcgttcc gaagggccat agcttcgagg agttggatgc cgttagccct
      421 cgcatgttgg gcgaagaaca agttctgctt ctgaccgagg gcaactgtat gcgtgaccag
      481 gtgcttagca tttttaccaa ggtcaaggtg cacatgaaga aagccctttc cggtgacagc
      541 tattgggtct ttgtgaaacg ggtaggtggc actggcggtg agcatgttgt ctttggcagt
      601 gaggacatcg agaacactat ggccaaaatg gacgacggac aactggatgg gttggccttt
      661 ggcgcaattc agctcgatgg tgacgggaat atcctgcagt acaatgctgc tgaaggagac
      721 atcacaggca gagatcccaa acaggtgatt gggaagaact tcttcaagga tgttgcacct
      781 ggaacggatt ctcccgagtt ttacggcaaa ttcaaggaag gcgtagcgtc agggaatctg
      841 aacaccatgt tcgaatggat gataccgact cttaccgatc atagttgcag cgagcttgcc
      901 gccaaacagc gcattcaggg tctcaccaac acccttcaag gctcaagcat cgataccatc
      961 cgccacatgg ttgccagtgg cttggcaatc agcgtcttgc ctgccactgc ccttaccgaa
     1021 aacgaccaca tgctgtttag cattatcccg tttgaaggca ccccgcctag ccgccgcgtt
     1081 gtcctggcct atcgccgcaa ctttgtccgc cctaaggccc tttcagccat gaaagccgca
     1141 attatgcaaa gccaactgca tggtgttagc ttcatctgcg atggggatcc accggtcatg
     1201 ggcgtggccg acttgatcaa gaagttcgag tccatctcca aggaggagtg actcgagcct
     1261 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     1321 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1381 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1441 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     1501 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     1561 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     1621 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1681 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1741 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1801 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1861 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1921 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1981 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2041 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2101 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2161 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2221 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2281 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2341 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2401 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2461 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2521 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2581 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2641 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2701 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2761 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2821 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     2881 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2941 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3001 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3061 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3121 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3181 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3241 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3301 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3361 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3421 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3481 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3541 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3601 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3661 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3721 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3781 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3841 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3901 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3961 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4021 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4081 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4141 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     4201 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaccg
     4261 ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg
     4321 ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag
     4381 ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc tggcactgtg
     4441 ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt
     4501 gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg
     4561 ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc tattggccag
     4621 gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg
     4681 acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc
     4741 gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg
     4801 tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa
     4861 atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat
     4921 gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt
     4981 aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact
     5041 attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg
     5101 cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa
     5161 tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca
     5221 atgctcgttt agggaac
//
