LOCUS       sequence-177729-        7035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Orange color variant of bright luminescent protein, fused to histone
            2B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7035)
  AUTHORS   Suzuki K, Kimura T, Shinoda H, Bai G, Daniels MJ, Arai Y, Nakano M, 
            Nagai T
  TITLE     Five colour variants of bright luminescent protein for real-time 
            multicolour bioimaging.
  JOURNAL   Nat Commun. 2016 Dec 14;7:13718. doi: 10.1038/ncomms13718.
  PUBMED    27966527
REFERENCE   2  (bases 1 to 7035)
  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: "Nat 
            Commun."; date: "2016-12-14"; volume: "7:13718. doi"; pages: " 
            10.1038/ncomms13718"
COMMENT     Sequence Label: OeNL-H2B_pcDNA3
FEATURES             Location/Qualifiers
     source          1..7035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        154..533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        534..737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     688..708
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     782..801
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        782..800
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1073..1723
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             1073..1723
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     promoter        complement(2507..2525)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2508..2525)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2545..2562)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2551..2775
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2821..3249
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2908..2927)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3118..3139
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3258..3278)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3263..3592
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3443..3578
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3505..3524
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3659..4453
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3713..3732)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4323..4342
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4627..4748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4664..4683)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4718..4737
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4797..4813)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4797..4813)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4810..4832)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4821..4837
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4845..4851,4852..4869,4870..4875))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4890..4911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5028..5045)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5199..5787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5279..5298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5958..6749,6750..6818))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6581..6600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6819..6923)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6998..7017)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
       61 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
      121 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
      181 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      241 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      301 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      361 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      421 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      481 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      541 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      601 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      661 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      721 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
      781 ttaatacgac tcactatagg gagacccaag cttggtaccg agctcggatc cgatggtctt
      841 cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt
      901 ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat
      961 ccaaaggatt gtcctgagcg gtctcgaggg cggtagcggt ggcagcgtga gcgtgatcaa
     1021 gcccgagatg aagaaggtgg aggacgccgt ggcccactcc actctcgagg gcgtgagcgt
     1081 gatcaagccc gagatgaaga tgaggtacta catggacggc tccgtcaatg ggcatgagtt
     1141 cacaatcgag ggtgagggca caggcagacc ttacgaggga catcaggaga tgacactgcg
     1201 cgtcacaatg gccgagggcg ggccaatgcc tttcgccttc gacctggtgt cccacgtgtt
     1261 ctgttacggc cacagagttt ttactaaata tccagaagag atcccagact atttcaagca
     1321 ggcctttcct gagggcctgt cctgggagag gtccctggag ttcgaggacg gcggctccgc
     1381 ctccgtgagc gcccacatca gcctgagggg caacaccttc taccacaagt ccaagttcac
     1441 cggcgtgaac ttccccgccg acggccccat catgcagaac cagagcgtgg actgggagcc
     1501 ctccaccgag aagatcaccg ccagcgacgg cgtgctgaag ggcgacgtga ccatgtacct
     1561 gaagctggag ggcggcggca accacaagtg ccagatgaag accacctaca aggccgccaa
     1621 ggagatcctg gagatgcccg gcgaccacta catcggccac aggctggtga ggaagaccga
     1681 gggcaacatc accgagcagg tggaggacgc cgtggcccac tccgagctcg aaaatgggct
     1741 gaagatcgac atccatgtca tcatcccgta tgaaggtctg agcggcgacc aaatgggcca
     1801 gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat catcacttta aggtgatcct
     1861 gcactatggc acactggtaa tcgacggggt tacgccgaac atgatcgact atttcggacg
     1921 gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc actgtaacag ggaccctgtg
     1981 gaacggcaac aaaattatcg acgagcgcct gatcaacccc gacggctccc tgctgttccg
     2041 agtaaccatc aacggagtga ccggctggcg gctgtgcgaa cgcattctgg cggaattcat
     2101 gccggagcca gcgaagtctg ctcccgcgcc caagaagggc tcgaagaaag ccgtgactaa
     2161 ggcgcagaag aaggacggca agaagcgcaa gcgcagccgc aaggagagct actccgtata
     2221 cgtgtacaag gtgctgaagc aggtccaccc cgacaccggc atctcctcta aggccatggg
     2281 aatcatgaac tccttcgtca acgacatctt cgaacgcatc gcgggtgagg cttcccgcct
     2341 ggcgcattac aacaagcgct cgaccatcac ctccagggag atccagacgg ccgtgcgcct
     2401 gctgctgccc ggggagttgg ccaagcacgc cgtgtccgag ggcaccaagg ccgtcaccaa
     2461 gtacaccagc gctaagtaac tcgagcatgc atctagaggg ccctattcta tagtgtcacc
     2521 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2581 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2641 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2701 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2761 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2821 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2881 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2941 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3001 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3061 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3121 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3181 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3241 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3301 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3361 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3421 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3481 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3541 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3601 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3661 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3721 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3781 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3841 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3901 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3961 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4021 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4081 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4141 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4201 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4261 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4321 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4381 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4441 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4501 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4561 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4621 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4681 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4741 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4801 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4861 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4921 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4981 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5041 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5101 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5161 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5221 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5281 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5341 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5401 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5461 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5521 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5581 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5641 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5701 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5761 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5821 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5881 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5941 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6001 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6061 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6121 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6181 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6241 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6301 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6361 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6421 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6481 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6541 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6601 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6661 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6721 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6781 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6841 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6901 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     6961 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     7021 agttaagcca gtatc
//
