LOCUS       sequence-116021-        5966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ONL-H2B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5966)
  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 5966)
  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_H2B
FEATURES             Location/Qualifiers
     source          1..5966
                     /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"
     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"
     polyA_signal    2754..2875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2791..2810)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2845..2864
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2882..3337)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3019..3040)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3231..3250
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3364..3468
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3470..3827
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3470..3661
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3493..3513)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3678..3813
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3862..4656
                     /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             3862..4656
                     /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(3916..3935)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4526..4545
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4844..4863)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4888..4935
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5264..5852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5753..5772
                     /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 ccatgccgga gccagcgaag tctgctcccg cgcccaagaa gggctcgaag aaagccgtga
     2281 ctaaggcgca gaagaaggac ggcaagaagc gcaagcgcag ccgcaaggag agctactccg
     2341 tatacgtgta caaggtgctg aagcaggtcc accccgacac cggcatctcc tctaaggcca
     2401 tgggaatcat gaactccttc gtcaacgaca tcttcgaacg catcgcgggt gaggcttccc
     2461 gcctggcgca ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc
     2521 gcctgctgct gcccggggag ttggccaagc acgccgtgtc cgagggcacc aaggccgtca
     2581 ccaagtacac cagcgctaag taagtcgacg gtaccgcggg cccgggatcc accggatcta
     2641 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2701 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2761 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2821 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     2881 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2941 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3001 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3061 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3121 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3181 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3241 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3301 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3361 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3421 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3481 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3661 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3721 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3781 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3841 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3901 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3961 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4021 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4081 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4141 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4201 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4261 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4321 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4381 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4441 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4501 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4561 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4621 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4681 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4741 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4801 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4861 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4921 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4981 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5041 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5101 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5161 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5221 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5281 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5341 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5401 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5461 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5521 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5581 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5641 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5701 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5761 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5821 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5881 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5941 tggataaccg tattaccgcc atgcat
//
