LOCUS       sequence-218251-        5566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pH sensor, targeted to mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5566)
  AUTHORS   Poburko D, Santo-Domingo J, Demaurex N
  TITLE     Dynamic regulation of the mitochondrial proton gradient during 
            cytosolic calcium elevations.
  JOURNAL   J Biol Chem. 2011 Apr 1;286(13):11672-84. doi: 
            10.1074/jbc.M110.159962. Epub 2011 Jan 11.
  PUBMED    21224385
REFERENCE   2  (bases 1 to 5566)
  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: 
            "10.1074/jbc.M110.159962"; journalName: "J Biol Chem"; date: 
            "2011-04-1- 1"; volume: "286"; issue: "13"; pages: "11672-84"
COMMENT     Sequence Label: SypHer mt
FEATURES             Location/Qualifiers
     source          1..5566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             651..725
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             747..818
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             852..2285
                     /codon_start=1
                     /product="improved fluorescent indicator for intracellular 
                     H2O2 (Bilan et al., 2013)"
                     /label=HyPer-3
                     /translation="EMASQQGETMSGPLHIGLIPTVGPYLLPHIIPMLHQTFPKLEMYL
                     HEAQTHQLLAQLDSGKLDCVILALVKESEAFIEVPLFDEPMLLAIYEDHPWANRECVPM
                     ADLAGEKLLMLEDGHSLRDQAMSAGYNSDNVYIMADKQKNGIKANFKIRHNVEDGSVQL
                     ADHYQQNTPIGDGPVLLPDNHYLSFQSVLSKDPNEKRDHMVLLEFVTAAGITLGMDELY
                     NVDGGSGGTGSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTT
                     GKLPVPWPTLVTTLGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTR
                     AEVKFEGDTLVNRIELKGIGFKEDGNILGHKLEYNGTGFCFEAGADEDTHFRATSLETL
                     RNMVAAGSGITLLPALAVPPERKRDGVVYLPCIKPEPRRTIGLVYRPGSPLRSRYEQLA
                     EAIRARMDGHFDKVLKQAV"
     primer_bind     1457..1478
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1587..1608)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1848..1867)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2408..2529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2445..2464)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2499..2518
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2536..2991)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2673..2694)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2885..2904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3018..3122
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3124..3481
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3124..3315
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3147..3167)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3332..3467
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3516..4310
                     /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             3516..4310
                     /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(3570..3589)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4180..4199
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4498..4517)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4542..4589
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4918..5506
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5407..5426
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc atgtccgtcc
      661 tgacgccgct gctgctgcgg ggcttgacag gctcggcccg gcggctccca gtgccgcgcg
      721 ccaagatcca ttcgttgggg gatctgtccg tcctgacgcc gctgctgctg cggggcttga
      781 caggctcggc ccggcggctc ccagtgccgc gcgccaagat ccattcgttg ggggatccac
      841 cggtcgccac cgagatggca agccagcagg gcgagacgat gtccggaccg ctgcacattg
      901 gtttgattcc cacagttgga ccgtacctgc taccgcatat tatccctatg ctgcaccaga
      961 cctttccaaa gctggaaatg tatctgcatg aagcacagac ccaccagtta ctggcgcaac
     1021 tggacagcgg caaactcgat tgcgtgatcc tcgcgctggt gaaagagagc gaagcattca
     1081 ttgaagtgcc gttgtttgat gagccaatgt tgctggctat ctatgaagat cacccgtggg
     1141 cgaaccgcga atgcgtaccg atggccgatc tggcagggga aaaactgctg atgctggaag
     1201 atggtcactc tttgcgcgat caggcaatgt ccgccggcta caacagcgac aacgtctata
     1261 tcatggccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacgtcg
     1321 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1381 ccgtgctgct gcccgacaac cactacctga gcttccagtc cgtcctgagc aaagacccca
     1441 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1501 gcatggacga gctgtacaac gtggatggcg gtagcggtgg caccggcagc aagggcgagg
     1561 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1621 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc
     1681 tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctcggct
     1741 acggcctgaa gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1801 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1861 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1921 agggcatcgg cttcaaggag gacggcaaca tcctggggca caagctggag tacaacggca
     1981 ccggtttctg ttttgaagcc ggggcggatg aagatacaca cttccgcgcg accagcctgg
     2041 aaactctgcg caacatggtg gcggcaggta gcgggatcac tttactgcca gcgctggctg
     2101 tgccgccgga gcgcaaacgc gatggggttg tttatctgcc gtgcattaag ccggaaccac
     2161 gccgcactat tggcctggtt tatcgtcctg gctcaccgct gcgcagccgc tatgagcagc
     2221 tggcagaggc catccgcgca agaatggatg gccatttcga taaagtttta aaacaggcgg
     2281 tttaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     2341 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2401 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2461 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2521 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2581 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2641 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2701 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2761 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2821 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2881 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2941 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3001 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3061 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3121 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3181 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3241 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3301 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3361 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3421 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3481 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3541 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3601 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3661 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3721 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3781 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3841 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3901 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3961 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4021 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4081 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4141 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4201 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4261 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4321 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4381 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4441 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4501 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4561 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4621 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4681 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4741 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4801 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4861 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4921 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4981 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5041 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5101 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5161 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5221 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5281 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5341 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5401 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5461 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5521 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatg
//
