LOCUS       sequence-271538-        5442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of F-actin targeted ultrasensitive hydrogen 
            peroxide indicator HyPer7 fused with LifeAct peptide for optical 
            imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5442)
  AUTHORS   Pak VV, Ezerina D, Lyublinskaya OG, Pedre B, Tyurin-Kuzmin PA, 
            Mishina NM, Thauvin M, Young D, Wahni K, Martinez Gache SA, 
            Demidovich AD, Ermakova YG, Maslova YD, Shokhina AG, Eroglu E, Bilan
            DS, Bogeski I, Michel T, Vriz S, Messens J, Belousov VV
  TITLE     Ultrasensitive Genetically Encoded Indicator for Hydrogen Peroxide 
            Identifies Roles for the Oxidant in Cell Migration and Mitochondrial
            Function.
  JOURNAL   Cell Metab. 2020 Mar 3;31(3):642-653.e6. doi: 
            10.1016/j.cmet.2020.02.003.
  PUBMED    32130885
REFERENCE   2  (bases 1 to 5442)
  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"; doi: 
            "10.1016/j.cmet.2020.02"; journalName: "Cell Metab"; date: 
            "2020-03-3- 3"; volume: "31"; issue: "3"; pages: "642-653.e6"
COMMENT     Sequence Label: pLifeAct-HyPer7
FEATURES             Location/Qualifiers
     source          1..5442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      233..242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             239..289
                     /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"
     primer_bind     916..937
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1046..1067)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1307..1326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    1866..1987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1903..1922)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1957..1976
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1994..2449)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2131..2152)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2343..2362
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2476..2580
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2582..2939
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2582..2773
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2605..2625)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2790..2925
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2974..3768
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2974..3768
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3028..3047)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3638..3657
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3956..3975)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4000..4047
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4376..4964
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4865..4884
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5139..5442
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgccaccat
      241 gggcgtggcc gacttgatca agaagttcga gtccatctcc aaggaggagg gggatccacc
      301 ggtcatgcac ctggctaatg aggagcaaaa cgaattggaa ggtgctttta agctgggctt
      361 gatctttacc gtcgccccgt acttgctgcc taagttgatc gtcagcctgc gccgcaccgc
      421 cccgaagatg cctttgatgt tggaggagaa ttacacccac acccttaccg agtcactgaa
      481 acgcggcgac gttgacgcca tcattgtggc agagccgttt caagagccgg gtattgtgac
      541 tgaacctttg tatgacgagc cgtttttcgt tatcgttccg aagggccata gcttcgagga
      601 gttggatgcc gttagccctc gcatgttggg cgaagaacaa gttctgcttc tgaccgaggg
      661 caactgtatg cgtgaccagg tgcttagctc cgccggcttc aacagcaaca acgtctatat
      721 catggccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacgtcga
      781 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      841 cgtgctgctg cccgacaacc actacctgag cttccagtcc gtcctgagca aagaccccaa
      901 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
      961 catggacgag ctgtacaacg tggatggcgg tagcggtggc accggcagca agggcgagga
     1021 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1081 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     1141 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctcagcta
     1201 cggcctgaag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1261 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1321 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1381 gggcatcggc ttcaaggagg acggcaacat cctggggcac aagctggagt acaacggccc
     1441 cagttgcagc gagcttgccg ccaaacagcg cattcagggt ctcaccaaca cccttcaagg
     1501 ctcaagcatc aataccatcc gccacatggt tgccagtggc ttggcaatca gcgtcttgcc
     1561 tgccactgcc cttaccgaaa acgaccacat gctgtttagc attatcccgt ttgaaggcac
     1621 cccgcctagc cgccgcgttg tcctggccta tcgccgcaac tttgtccgcc ctaaggccct
     1681 ttcagccatg aaagccgcaa ttatgcaaag ccaactgcat ggtgttagct tcatctgcga
     1741 ttgagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     1801 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     1861 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1921 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1981 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2041 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2101 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2161 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2221 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2281 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2341 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2401 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     2461 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2521 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2581 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2701 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2761 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2821 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2881 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     2941 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3001 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3061 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3121 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     3181 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3241 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3301 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3361 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3421 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3481 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     3541 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3601 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3661 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3721 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     3781 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     3841 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3901 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     3961 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4021 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4081 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     4141 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     4201 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     4261 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4321 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4381 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4441 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4501 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4561 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4621 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4681 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4741 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4801 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4861 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4921 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4981 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5041 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
     5101 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     5161 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     5221 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     5281 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     5341 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     5401 acttggcagt acatctacgt attagtcatc gctattacca tg
//
