LOCUS       sequence-317302-        5559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mitochondria-targeted genetically encoded hydrogen peroxide 
            indicator with red fluorescence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5559)
  AUTHORS   Ermakova YG, Bilan DS, Matlashov ME, Mishina NM, Markvicheva KN, 
            Subach OM, Subach FV, Bogeski I, Hoth M, Enikolopov G, Belousov VV
  TITLE     Red fluorescent genetically encoded indicator for intracellular 
            hydrogen peroxide.
  JOURNAL   Nat Commun. 2014 Oct 21;5:5222. doi: 10.1038/ncomms6222.
  PUBMED    25330925
REFERENCE   2  (bases 1 to 5559)
  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 
            Commun."; date: "2014-10-21"; volume: "5:5222. doi"; pages: " 
            10.1038/ncomms6222"
COMMENT     Sequence Label: pC1-HyPerRed-mito
FEATURES             Location/Qualifiers
     source          1..5559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             538..612
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             634..705
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             join(739..1116,1117..1125,1126..1851,1852..1860,1861..2163)
                     /codon_start=1
                     /product="red fluorescent genetically encoded indicator for
                     H2O2 ()"
                     /label=HyPerRed
                     /translation="EMASQQGETMSGPLHIGLIPTVGPYLLPHIIPMLHQTFPKLEMYL
                     HEAQTHQLLAQLDSGKLDCVILALVKESEAFIEVPLFDEPMLLAIYEDHPWANRECVPM
                     ADLAGEKLLMLEDGHCLRDQAMPRTVSERMYPEDGALKSEIKKGLRLKDGGHYAAEVKT
                     TYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQCERAEGRHSTGGMDELYKGGTGGSL
                     VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEAFQTAKLKVTKGGPLP
                     FAWDILSPQFMYGSKAYIKHPADIPDYFKLSFPEGFRWERVMNFEDGGIIHVNQDSSLQ
                     DGVFIYKVKLRGTNFPPDGPVMQKKTMGWEAAGVGFCFEAGADEDTHFRATSLETLRNM
                     VAAGSGITLLPALAVPPERKRDGVVYLPCIKPEPRRTIGLVYRPGSPLRSRYEQLAEAI
                     RARMDGHFDKVLKQAV"
     primer_bind     1815..1834
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2288..2409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2325..2344)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2379..2398
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2416..2871)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2553..2574)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2765..2784
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2898..3002
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3004..3361
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3004..3195
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3027..3047)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3212..3347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3396..4190
                     /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             3396..4190
                     /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(3450..3469)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4060..4079
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4378..4397)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4422..4469
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4798..5386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5287..5306
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcatg
      541 tccgtcctga cgccgctgct gctgcggggc ttgacaggct cggcccggcg gctcccagtg
      601 ccgcgcgcca agatccattc gttgggggat ctgtccgtcc tgacgccgct gctgctgcgg
      661 ggcttgacag gctcggcccg gcggctccca gtgccgcgcg ccaagatcca ttcgttgggg
      721 gatccaccgg tcgccaccga gatggcgagc cagcagggcg agacgatgtc cggaccgctg
      781 cacattggtt tgattcccac agttggaccg tacctgctac cgcatattat ccctatgctg
      841 caccagacct ttccaaagct ggaaatgtat ctgcatgaag cacagaccca ccagttactg
      901 gcgcaactgg acagcggcaa actcgattgc gtgatcctcg cactggtgaa agagagcgaa
      961 gcattcattg aagtgccgtt gtttgatgag ccaatgttgc tggctatcta tgaagatcac
     1021 ccgtgggcga accgcgaatg cgtaccgatg gccgatctgg caggggaaaa actgctgatg
     1081 ctggaagatg gtcactgttt gcgcgatcag gcaatgcccc gcacggtttc cgagcggatg
     1141 taccccgagg acggcgccct gaagagcgag atcaagaagg ggctgaggct gaaggacggc
     1201 ggccactacg ccgccgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     1261 ggcgcctaca tcgtcgacat caagttggac atcgtgtccc acaacgagga ctacaccatc
     1321 gtggaacagt gcgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     1381 aagggaggta caggcgggag tctggtgagc aagggcgagg aggataacat ggccatcatc
     1441 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     1501 atcgagggcg agggcgaggg ccgcccctac gaggcctttc agaccgctaa gctgaaggtg
     1561 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     1621 tccaaggcct acattaagca cccagccgac atccccgact acttcaagct gtccttcccc
     1681 gagggcttca ggtgggagcg cgtgatgaac ttcgaggacg gcggcattat tcacgttaac
     1741 caggactcct ccctgcagga cggcgtattc atctacaagg tgaagctgcg cggcaccaac
     1801 ttcccccccg acggccccgt aatgcagaag aagaccatgg gctgggaggc tgccggggtg
     1861 ggtttctgtt ttgaagccgg ggcggatgaa gatacacact tccgcgcgac cagcctggaa
     1921 actctgcgca acatggtggc ggcaggtagc gggatcactt tactgccagc gctggctgtg
     1981 ccgccggagc gcaaacgcga tggggttgtt tatctgccgt gcattaagcc ggaaccacgc
     2041 cgcactattg gtctggttta tcgtcctggc tcaccgctgc gcagccgcta tgagcagctg
     2101 gcagaggcca tccgcgcaag aatggatggc catttcgata aagttttaaa acaggcggtt
     2161 taatgagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     2221 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2281 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2341 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2401 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2461 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2521 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2581 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2641 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2701 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2761 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2821 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     2881 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2941 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3001 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3061 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3121 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3181 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3241 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3301 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3361 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3421 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3481 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3541 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3601 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3661 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3721 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3781 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3841 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3901 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     3961 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4021 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4081 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4141 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4201 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4261 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4321 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4381 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4441 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4501 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4561 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4621 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4681 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4741 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4801 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4861 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4921 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4981 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5041 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5101 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5161 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5221 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5281 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5341 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5401 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5461 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     5521 ttacggggtc attagttcat agcccatata tggagttcc
//
