LOCUS       sequence-271540-        5720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mitochondrial matrix targeted ultrasensitive
            hydrogen peroxide indicator HyPer7 for optical imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5720)
  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 5720)
  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: pCS2+MLS-HyPer7
FEATURES             Location/Qualifiers
     source          1..5720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1084..1158
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1180..1251
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     1884..1905
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2014..2035)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2275..2294)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2750..2884
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2774..2793)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2828..2847
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2997..3017)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2997..3015)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3036..3052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3036..3052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3049..3071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3060..3076
                     /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(3084..3090,3091..3108,3109..3114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3129..3150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3267..3284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3438..4026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3518..3537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4197..4988,4989..5057))
                     /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     4820..4839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5058..5162)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5188..5643)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5325..5346)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5537..5556
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt cgaattcgcc
     1081 accatgtccg tcctgacgcc gctgctgctg cggggcttga caggctcggc ccggcggctc
     1141 ccagtgccgc gcgccaagat ccattcgttg ggggatctgt ccgtcctgac gccgctgctg
     1201 ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc cgcgcgccaa gatccattcg
     1261 ttgggggata ccggtcacct ggctaatgag gagcaaaacg aattggaagg tgcttttaag
     1321 ctgggcttga tctttaccgt cgccccgtac ttgctgccta agttgatcgt cagcctgcgc
     1381 cgcaccgccc cgaagatgcc tttgatgttg gaggagaatt acacccacac ccttaccgag
     1441 tcactgaaac gcggcgacgt tgacgccatc attgtggcag agccgtttca agagccgggt
     1501 attgtgactg aacctttgta tgacgagccg tttttcgtta tcgttccgaa gggccatagc
     1561 ttcgaggagt tggatgccgt tagccctcgc atgttgggcg aagaacaagt tctgcttctg
     1621 accgagggca actgtatgcg tgaccaggtg cttagctccg ccggcttcaa cagcaacaac
     1681 gtctatatca tggccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1741 aacgtcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1801 gacggccccg tgctgctgcc cgacaaccac tacctgagct tccagtccgt cctgagcaaa
     1861 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1921 actctcggca tggacgagct gtacaacgtg gatggcggta gcggtggcac cggcagcaag
     1981 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2041 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2101 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2161 ctcagctacg gcctgaagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     2221 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2281 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2341 gagctgaagg gcatcggctt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2401 aacggcccca gttgcagcga gcttgccgcc aaacagcgca ttcagggtct caccaacacc
     2461 cttcaaggct caagcatcaa taccatccgc cacatggttg ccagtggctt ggcaatcagc
     2521 gtcttgcctg ccactgccct taccgaaaac gaccacatgc tgtttagcat tatcccgttt
     2581 gaaggcaccc cgcctagccg ccgcgttgtc ctggcctatc gccgcaactt tgtccgccct
     2641 aaggcccttt cagccatgaa agccgcaatt atgcaaagcc aactgcatgg tgttagcttc
     2701 atctgcgatt gactcgagcc tctagaacta tagtgagtcg tattacgtag atccagacat
     2761 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2821 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2881 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2941 tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     3001 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3061 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     3121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3841 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3961 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4021 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4081 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4141 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4201 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4261 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4321 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4381 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4441 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4501 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4561 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4621 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4681 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4741 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4801 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4861 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4921 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4981 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5041 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5101 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5161 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     5221 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     5281 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     5341 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     5401 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     5461 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     5521 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     5581 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     5641 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     5701 cgctattacg ccagtcgacc
//
