LOCUS       sequence-433842-        5764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting of the chemigenetic, fluorescent peroxide sensor oROS-HT 
            to mitochondrial matrix..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5764)
  AUTHORS   Lee J.D., Nguyen A., Jin Z.R., Moghadasi A., Gibbs C.E., Wait S.J., 
            Evitts K.M., Asencio A., Bremner S.B., Zuniga S., Chavan V., 
            Williams A., Smith N., Regnier M., Young J.E., Mack D., Nance E., 
            Boyle P.M., Berndt A.
  TITLE     Far-red and sensitive sensor for monitoring real time H2O2 dynamics 
            with subcellular resolution and in multi-parametric imaging 
            applications
  JOURNAL   bioRxiv 2024.02.06.579232
REFERENCE   2  (bases 1 to 5764)
  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: "bioRxiv 
            2024.02.06.579232"
COMMENT     Sequence Label: pC1-dmito-oROS-HT
FEATURES             Location/Qualifiers
     source          1..5764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    123..244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(160..179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     214..233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(251..706)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(388..409)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     600..619
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        733..837
                     /gene="bla"
                     /label=AmpR promoter
     promoter        839..1196
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        839..1030
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(862..882)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1047..1182
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1231..2025
                     /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             1231..2025
                     /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(1285..1304)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1895..1914
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2213..2232)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2257..2304
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2633..3221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3122..3141
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3396..3699
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3700..3903
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3854..3874
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3932..4006
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             4028..4099
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
ORIGIN
        1 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
       61 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
      121 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
      181 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
      241 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      301 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      361 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      421 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      481 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      541 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      601 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      661 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
      721 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      781 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
      841 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
      901 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
      961 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1021 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     1081 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     1141 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     1201 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     1261 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     1321 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     1381 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     1441 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     1501 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     1561 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     1621 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     1681 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     1741 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     1801 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     1861 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     1921 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     1981 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     2041 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     2101 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     2161 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     2221 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     2281 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     2341 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     2401 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     2461 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     2521 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2581 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2641 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2701 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2761 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2821 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2881 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2941 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3001 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3061 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3121 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3181 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3241 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3301 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     3361 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     3421 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     3481 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     3541 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     3601 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     3661 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     3721 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     3781 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     3841 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     3901 gctggtttag tgaaccgtca gatccgctag catgtccgtc ctgacgccgc tgctgctgcg
     3961 gggcttgaca ggctcggccc ggcggctccc agtgccgcgc gccaagatcc attcgttggg
     4021 ggatctgtcc gtcctgacgc cgctgctgct gcggggcttg acaggctcgg cccggcggct
     4081 cccagtgccg cgcgccaaga tccattcgtt gggggatcca atggagatgg cgagccagca
     4141 gggcgagacg atgtccggac cgctgcacat tggtttgatt cccacagttg gaccgtacct
     4201 gctaccgcat attatcccta tgctgcacca gacctttcca aagctggaaa tgtatctgca
     4261 tgaagcacag acccaccagt tactggcgca actggacagc ggcaaactcg attgcgtgat
     4321 cctcgcactg gtgaaagaga gcgaagcatt cattgaagtg ccgttgtttg atgagccaat
     4381 gttgctggct atctatgaag atcacccgtg ggcgaaccgc gaatgcgtac cgatggccga
     4441 tctggcaggg gaaaaactgc tgatgctgga agatggtcac tgtttgcgcg atcaggcaat
     4501 gggtttctgt tttgaagccg gggcgctgag ctcaacgggt gagaccttcc aggccttccg
     4561 caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct
     4621 gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt
     4681 cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg
     4741 tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc
     4801 tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc
     4861 tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct
     4921 gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtcgacgct
     4981 cgagatttcc ggcggaggaa caggtggttc tggtggaaca gggggtagcg gaggtacagg
     5041 aggaagtatg gcggagatcg gaactggatt cccgtttgat ccgcattatg tggaagttct
     5101 gggagagcgc atgcattatg tggacgttgg tcctcgtgat gggacaccag tgctgttcct
     5161 tcacggcaat ccgacatcgt cgtacgtgtg gcgtaatatc atcccgcacg ttgcccccac
     5221 gcaccgctgc attgcccctg acttaattgg tatggggaaa agtgataagc ctgatctggg
     5281 gtacttcttt gacgaccacg tacgcttcat ggatgctttt attgaagcat tgggtttgga
     5341 ggaagtagtt ttggtgatcc atgattgggg tagtgctctg gggttccatt gggccaagcg
     5401 taacccagaa cgcgtgaaag gaattgcctt tatggagttc atccgtccga ttccaacatg
     5461 ggacgaatgg cgaccggacc aactggatga agatacacac ttccgcgcga ccagcctgga
     5521 aactctgcgc aacatggtgg cggcaggtag cgggatcact ttactgccag cgctggctgt
     5581 gccgccggag cgcaaacgcg atggggttgt ttatctgccg tgcattaagc cggaaccacg
     5641 ccgcactatt ggtctggttt atcgtcctgg ctcaccgctg cgcagccgct atgagcagct
     5701 ggcagaggcc atccgcgcaa gaatggatgg ccatttcgat aaagttttaa aacaggcggt
     5761 ttaa
//
