LOCUS       sequence-111684-        5588 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses ONL-fibrillarin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5588)
  AUTHORS   Takai A, Nakano M, Saito K, Haruno R, Watanabe TM, Ohyanagi T, Jin 
            T, Okada Y, Nagai T
  TITLE     Expanded palette of Nano-lanterns for real-time multicolor 
            luminescence imaging.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):4352-6. doi: 
            10.1073/pnas.1418468112. Epub 2015 Mar 23.
  PUBMED    25831507
REFERENCE   2  (bases 1 to 5588)
  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: 
            "10.1073/pnas.1418468112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-04-7- 7"; volume: "112"; issue: "14"; pages: "4352-6"
COMMENT     Sequence Label: pONL-C1_fibrillarin
FEATURES             Location/Qualifiers
     source          1..5588
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     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"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             603..1256
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             603..1256
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     misc_feature    2187..2252
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2376..2497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2413..2432)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2467..2486
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2504..2959)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2641..2662)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2853..2872
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2986..3090
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3092..3449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3092..3283
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3115..3135)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3300..3435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3484..4278
                     /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             3484..4278
                     /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(3538..3557)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4148..4167
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4466..4485)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4510..4557
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4886..5474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5375..5394
                     /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 gctagcgcca
      601 ccatggtgag cgtgatcaag cccgagatga agatgaggta ctacatggac ggctccgtca
      661 atgggcatga gttcacaatc gagggtgagg gcacaggcag accttacgag ggacatcagg
      721 agatgacact gcgcgtcaca atggccgagg gcgggccaat gcctttcgcc ttcgacctgg
      781 tgtcccacgt gttctgttac ggccacagag tttttactaa atatccagaa gagatcccag
      841 actatttcaa gcaggccttt cctgagggcc tgtcctggga gaggtccctg gagttcgagg
      901 acggcggctc cgcctccgtg agcgcccaca tcagcctgag gggcaacacc ttctaccaca
      961 agtccaagtt caccggcgtg aacttccccg ccgacggccc catcatgcag aaccagagcg
     1021 tggactggga gccctccacc gagaagatca ccgccagcga cggcgtgctg aagggcgacg
     1081 tgaccatgta cctgaagctg gagggcggcg gcaaccacaa gtgccagatg aagaccacct
     1141 acaaggccgc caaggagatc ctggagatgc ccggcgacca ctacatcggc cacaggctgg
     1201 tgaggaagac cgagggcaac atcaccgagc aggtggagga cgccgtggcc cactccggta
     1261 ccaaggtgta cgaccccgag caacgcaaac gcatgatcac tgggcctcag tggtgggctc
     1321 gctgcaagca aatgaacgtg ctggactcct tcatcaacta ctatgattcc gagaagcacg
     1381 ccgagaacgc cgtgattttt ctgcatggta acgctacctc cagctacctg tggaggcacg
     1441 tcgtgcctca catcgagccc gtggctagat gcatcatccc tgatctgatc ggaatgggta
     1501 agtccggcaa gagcgggaat ggctcatatc gcctcctgga tcactacaag tacctcaccg
     1561 cttggttcga gctgctgaac cttccaaaga aaatcatctt tgtgggccac gactggggga
     1621 gcgctctggc ctttcactac gcctacgagc accaagacag gatcaaggcc atcgtccata
     1681 tggagagtgt cgtggacgtg atcgagtcct ggatggggtg gcctgacatc gaggaggagc
     1741 tggccctgat caagagcgaa gagggcgaga aaatggtgct tgagaataac ttcttcgtcg
     1801 agaccctgtt gccaagcaag atcatgcgga aactggagcc tgaggagttc gctgcctacc
     1861 tggagccatt caaggagaag ggcgaggtta gacggcctac cctctcctgg cctcgcgaga
     1921 tccctctcgt taagggaggc aagcccgacg tcgtccagat tgtccgcaac tacaacgcct
     1981 accttcgggc cagcgacgat ctgcctaagc tgttcatcga gtccgaccct gggttctttt
     2041 ccaacgctat tgtcgaggga gctaagaagt tccctaacac cgagttcgtg aaggtgaagg
     2101 gcctccactt cctccaggag gacgctccag atgaaatggg taagtacatc aagagcttcg
     2161 tggagcgcgt gctgaagaac gagcagtccg gactcagatc tcgagctcaa gcttcgaatt
     2221 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     2281 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2341 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2401 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2461 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     2521 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2581 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2641 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2701 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2761 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2821 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2881 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2941 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3001 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3061 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3121 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3181 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3241 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3301 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3361 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3421 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3481 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3541 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3601 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3661 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3721 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3781 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3841 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3901 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3961 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4021 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4081 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4141 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4201 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4261 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4321 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4381 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4441 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4501 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4561 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4621 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4681 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4741 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4801 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4861 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4921 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4981 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     5041 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5101 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5161 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5221 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5281 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5341 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5401 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5461 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5521 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5581 ccatgcat
//
