LOCUS       sequence-435274-        5764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting of the chemigenetic, fluorescent peroxide sensor oROS-HT 
            to the inter-membrane-space of mitochondria..
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-IMS-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
     primer_bind     complement(862..882)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      996..1131
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1180..1974
                     /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             1180..1974
                     /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(1234..1253)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1844..1863
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2162..2181)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2206..2253
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2582..3170
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3071..3090
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3345..3648
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3649..3852
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3803..3823
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3881..3890
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3885..3894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4115..4124
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
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 atagtcccgc
      961 ccctaactcc gcccatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     1021 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     1081 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     1141 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     1201 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     1261 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     1321 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     1381 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     1441 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     1501 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     1561 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     1621 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     1681 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     1741 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     1801 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     1861 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     1921 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     1981 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     2041 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     2101 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     2161 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     2221 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     2281 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     2341 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     2401 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     2461 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2521 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2581 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2641 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2701 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2761 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2821 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2881 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2941 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3001 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3061 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3121 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3181 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3241 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
     3301 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     3361 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     3421 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     3481 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     3541 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     3601 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     3661 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     3721 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     3781 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     3841 ataagcagag ctggtttagt gaaccgtcag atccgctagc gccaccatgg cggctctgaa
     3901 gagttggctg tcgcgcagcg taacttcatt cttcaggtac agacagtgtt tgtgtgttcc
     3961 tgttgtggct aactttaaga agcggtgttt ctcagaattg ataagaccat ggcacaaaac
     4021 tgtgacgatt ggctttggag taaccctgtg tgcggttcct attaagcttc gaattctgca
     4081 gtcgacggta ccgcgggccc gggatccacc cgtcgccacc 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
//
