LOCUS       sequence-330582-        6862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chemogenetic fluorescent H2O2 reporter targeted to the mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6862)
  AUTHORS   Saeedi Saravi SS, Eroglu E, Waldeck-Weiermair M, Sorrentino A, 
            Steinhorn B, Belousov V, Michel T
  TITLE     Differential endothelial signaling responses elicited by 
            chemogenetic H2O2 synthesis.
  JOURNAL   Redox Biol. 2020 Sep;36:101605. doi: 10.1016/j.redox.2020.101605. 
            Epub 2020 Jun 16.
  PUBMED    32590330
REFERENCE   2  (bases 1 to 6862)
  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: "Redox 
            Biol."; date: "2020-09"; volume: "36:101605. doi"; pages: " 
            10.1016/j.redox.2020.101605. Epub 2020 Jun 16"
COMMENT     Sequence Label: HyPer7.2DAAO-mito
FEATURES             Location/Qualifiers
     source          1..6862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     89..108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     342..359
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    476..497
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(512..517,518..535,536..542)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    550..566
                     /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)."
     primer_bind     555..577
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     574..590
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     574..590
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     609..629
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        611..629
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    742..876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(779..798)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     833..852
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(925..951)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     2474..2493
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2733..2754
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2863..2884)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(3517..3588)
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             complement(3610..3684)
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     promoter        complement(3737..3755)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3738..3755)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(3790..4774)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      4845..5300
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4932..4951)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5142..5163
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5326..5430
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5431..5499,5500..6291)
                     /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     complement(5649..5668)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6462..6862,1..188)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
       61 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      121 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      181 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      241 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      301 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      361 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      421 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      481 tgagttagct cactcattag gcaccccagg ctttacactt tatgctcccg gctcgtatgt
      541 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
      601 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct gggtaccggg cccaatgcat
      661 tggcgccgcg gccgcgaatt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
      721 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
      781 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
      841 tccaaactca tcaatgtatc ttatcatgtc tggatctacg taatacgact cactatagtt
      901 ctagaggctc gaggcaagct tttacagggt cagccgctcc agggggggca ggctctccct
      961 agctgcgccg tggtatcttt ggaaggcttc atcgaccagc tgagccacgt cctccacggc
     1021 tccccaggat tgctgatacc ctgcgctact aaagccgtag gcgtggacaa gggtcacttc
     1081 cttctccatt gcagcccgtg cggaccctct tcccagagag agagggctct tggtccggtc
     1141 caggggcaag acgattcttt cagcctccac cctaggtcct cctctcctgg cgggtctcag
     1201 gccaacgtta tgtctcagca cttcaatgcc ctcgatagtt ccatcggagc taatggtagg
     1261 gtcaagtctc aggcagtgct tgagaatcct ttggactgtt tcgggattca cgctcaggtc
     1321 ccaatcgccg actccgtagg ttccgccaca gatcacttct cctcctggtc taggaatgat
     1381 ataagcggga gaagcagggt cggagctatc cattgtgcac ctcttacagg gagatttgac
     1441 aagcacggtc tgtcccctaa tgggctcggc agcctggtca tcgatgccgg caatggactt
     1501 agctcccagc ccggtggcgt tgaccacgag gtcagctcca tcgaaggctt gctccaggct
     1561 agtcactgtt ctcctttcga aggtagcccc aagtttctgc agctctctgg caaggtatct
     1621 gcaatacttg ggagcgtgga cgctcagggt gtcgtatgtc actccaatgg ctccaggtgg
     1681 acattcagaa ggtggaagtg gcctatagtt aggggtgatg tccttgtacc aatgtcccag
     1741 gaggccatcc tcattctggg caaaccttct ggtccctttc agccacatag cgtggcctgt
     1801 gggaaccagc tccacccatt tcttgaaggt ggactcttcc cacttggctt gtctgggtcc
     1861 gtctgtcagg gtcataaatg gtgtccagtt agctccggcc caggggcttg cgaaggtctg
     1921 ggagctgaca tcctcaggca agtctctggc caggatatgc acggagtagc cttttctagc
     1981 gagaatcagg gcgccagaga gcccgatgac tccggagccc agcacgacca ccctcttctg
     2041 gctgtgacca ctgctagcat cgcagatgaa gctaacacca tgcagttggc tttgcataat
     2101 tgcggctttc atggctgaaa gggccttagg gcggacaaag ttgcggcgat aggccaggac
     2161 aacgcggcgg ctaggcgggg tgccttcaaa cgggataatg ctaaacagca tgtggtcgtt
     2221 ttcggtaagg gcagtggcag gcaagacgct gattgccaag ccactggcaa ccatgtggcg
     2281 gatggtattg atgcttgagc cttgaagggt gttggtgaga ccctgaatgc gctgtttggc
     2341 ggcaagctcg ctgcaactgg ggccgttgta ctccagcttg tgccccagga tgttgccgtc
     2401 ctccttgaag ccgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     2461 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     2521 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     2581 ggcgaagcac ttcaggccgt agctgagggt ggtcacgagg gtgggccagg gcacgggcag
     2641 cttgccggtg gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc
     2701 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     2761 caccaccccg gtgaacagct cctcgccctt gctgccggtg ccaccgctac cgccatccac
     2821 gttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     2881 catgtgatcg cgcttctcgt tggggtcttt gctcaggacg gactggaagc tcaggtagtg
     2941 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     3001 gagctgcacg ctgccgtcct cgacgttgtg gcggatcttg aagttggcct tgatgccgtt
     3061 cttctgcttg tcggccatga tatagacgtt gttgctgttg aagccggcgg agctaagcac
     3121 ctggtcacgc atacagttgc cctcggtcag aagcagaact tgttcttcgc ccaacatgcg
     3181 agggctaacg gcatccaact cctcgaagct atggcccttc ggaacgataa cgaaaaacgg
     3241 ctcgtcatac aaaggttcag tcacaatacc cggctcttga aacggctctg ccacaatgat
     3301 ggcgtcaacg tcgccgcgtt tcagtgactc ggtaagggtg tgggtgtaat tctcctccaa
     3361 catcaaaggc atcttcgggg cggtgcggcg caggctgacg atcaacttag gcagcaagta
     3421 cggggcgacg gtaaagatca agcccagctt aaaagcacct tccaattcgt tttgctcctc
     3481 attagccagg tgaccggtat cccccaacga atggatcttg gcgcgcggca ctgggagccg
     3541 ccgggccgag cctgtcaagc cccgcagcag cagcggcgtc aggacggaca gatcccccaa
     3601 cgaatggatc ttggcgcgcg gcactgggag ccgccgggcc gagcctgtca agccccgcag
     3661 cagcagcggc gtcaggacgg acatggtggc gaattcgaat cgatgggatc ctgcaaaaag
     3721 aacaagtagc ttgtattcta tagtgtcacc taaatcaagc ttgctccgac gtccccaggc
     3781 agaatggcgg ttccctaaac gagcattgct tatatagacc tcccattagg cacgcctacc
     3841 gcccatttac gtcaatggaa cgcccatttg cgtcattgcc cctccccatt gacgtcaatg
     3901 gggatgtact tggcagccat cgcgggccat ttaccgccat tgacgtcaat gggagtactg
     3961 ccaatgtacc ctggcgtact tccaatagta atgtacttgc caagttacta ttaatagata
     4021 ttgatgtact gccaagtggg ccatttaccg tcattgacgt caataggggg cgtgagaacg
     4081 gatatgaatg ggcaatgagc catcccattg acgtcaatgg tgggtggtcc tattgacgtc
     4141 aatgggcatt gagccaggcg ggccatttac cgtaattgac gtcaatgggg gaggcgccat
     4201 atacgtcaat aggaccgccc atatgacgtc aataggaaag accatatata gagaccattg
     4261 acgtcaatgg gggagtggct atgggcggta ttaggaagcc ccatatatgg tatatgggac
     4321 cgcccattgg gaggggctat ctacgtcaat aggaaaaccc atatatggaa tactatatgg
     4381 catagggcca atacatagta ttgaacctgg ccaatagcca tattggcata gggccatatt
     4441 ggatattgcc tatatattga tcctggcata tagccaatat ggccgccatt attggcacca
     4501 tgccaattca atatggcgga cttggcacag ggccaagacc cctcctcaaa cttggcaccg
     4561 tgccaagtag acccctcccc agttggcaca gtgccaggtc cgccatattg aattggcatg
     4621 gtgccaagtc cgccatattg aattggctgg ggtccagatc caccatattg aattggcacg
     4681 agtccatata cgccatattg aattggcaca ggtccagatc caccatattg aattggcatg
     4741 agtccatata cgccatattg aattggcggt cgactggcgt aatagcgaag aggcccgcac
     4801 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     4861 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4921 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4981 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5041 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5101 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5161 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     5221 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5281 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5341 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5401 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5461 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5521 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5581 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5641 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5701 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5761 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5821 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5881 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5941 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6001 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6061 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6121 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6181 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6241 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6301 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6361 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6421 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6481 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6541 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6601 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6661 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6721 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6781 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6841 ctacagcgtg agctatgaga aa
//
