LOCUS       sequence-258765-        4846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SuperNova fused to mitochrondrial protein COX8A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4846)
  AUTHORS   Paardekooper LM, van Vroonhoven E, Ter Beest M, van den Bogaart G
  TITLE     Radical Stress Is More Cytotoxic in the Nucleus than in Other 
            Organelles.
  JOURNAL   Int J Mol Sci. 2019 Aug 25;20(17). pii: ijms20174147. doi: 
            10.3390/ijms20174147.
  PUBMED    31450682
REFERENCE   2  (bases 1 to 4846)
  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: "Int J Mol 
            Sci."; date: "2019-08-25"; volume: "20(17). pii: ijms20174147. doi";
            pages: " 10.3390/ijms20174147"
COMMENT     Sequence Label: COX8-SuperNova
FEATURES             Location/Qualifiers
     source          1..4846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             619..693
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      757..766
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(763..765,766..771,772..1479)
                     /codon_start=1
                     /product="monomeric variant of the photosensitizing red 
                     fluorescent protein KillerRed (Takemoto et al., 2013)"
                     /label=SuperNova
                     /translation="MGSEVGPALFQSDMTFKIFIDGEVNGQKFTIVADGSSKFPHGDFN
                     VHAVCETGKLPMSWKPICHLIQYGEPFFARYPDGISHFAQECFPEGLSIDRTVRFENDG
                     TMTSHHTYELDDTCVVSRITVNCDGFQPDGPIMRDQLVDILPSETHMFPHGPNAVRQTA
                     TIGFTTADGGKMMGHFDSKMTFNGSRAIEIPGPHFVTIITKQTRDTSDKRDHVCQREVA
                     YAHSVPRITSAIGSDED"
     CDS             1480..1509
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1634..1755
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1671..1690)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1725..1744
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1762..2217)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1899..1920)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2111..2130
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2244..2348
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2350..2707
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2350..2541
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2373..2393)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2558..2693
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2742..3536
                     /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             2742..3536
                     /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(2796..2815)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3406..3425
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3724..3743)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3768..3815
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4144..4732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4633..4652
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagat gagcgtgctg acccccctgc tgctgcgcgg cctgaccggc
      661 agcgcccgcc gcctgcccgt gccccgcgcc aagatccaca gcctgaagct tcgaattctg
      721 cagtcgacgg taccgcgggc ccgggatcca cccgtggcca ccatgggcag cgaggtgggc
      781 cccgccctgt tccagagcga catgaccttc aagatcttca tcgacggcga ggtgaacggc
      841 cagaagttca ccatcgtggc cgacggcagc agcaagttcc cccacggcga cttcaacgtg
      901 cacgccgtgt gcgagaccgg caagctgccc atgagctgga agcccatctg ccacctgatc
      961 cagtacggcg agcccttctt cgcccgctac cccgacggca tcagccactt cgcccaggag
     1021 tgcttccccg agggcctgag catcgaccgc accgtgcgct tcgagaacga cggcaccatg
     1081 accagccacc acacctacga gctggacgac acctgcgtgg tgagccgcat caccgtgaac
     1141 tgcgacggct tccagcccga cggccccatc atgcgcgacc agctggtgga catcctgccc
     1201 agcgagaccc acatgttccc ccacggcccc aacgccgtgc gccagaccgc caccatcggc
     1261 ttcaccaccg ccgacggcgg caagatgatg ggccacttcg acagcaagat gaccttcaac
     1321 ggcagccgcg ccatcgagat ccccggcccc cacttcgtga ccatcatcac caagcagacc
     1381 cgcgacacca gcgacaagcg cgaccacgtg tgccagcgcg aggtggccta cgcccacagc
     1441 gtgccccgca tcaccagcgc catcggcagc gacgaggacg agcagaagct gatcagcgag
     1501 gaggacctgt aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     1561 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     1621 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1681 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1741 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1801 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1861 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1921 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1981 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2041 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2101 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2161 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     2221 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2281 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2341 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     2401 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2461 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2521 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2581 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2641 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2701 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     2761 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2821 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     2881 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     2941 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3001 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3061 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3121 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3181 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3241 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     3301 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3361 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3421 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3481 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3541 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3601 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3661 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     3721 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     3781 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     3841 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     3901 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     3961 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     4021 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4081 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4141 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4201 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4261 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4321 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4381 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4441 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4501 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4561 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4621 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4681 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4741 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4801 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
