LOCUS       sequence-177725-        7860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Orange color variant of bright luminescent protein, fused to actin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7860)
  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 7860)
  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-Actin_pcDNA3
FEATURES             Location/Qualifiers
     source          1..7860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        164..543
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        544..747
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     698..718
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     792..811
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        792..810
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      825..834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1071..1721
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             1071..1721
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     promoter        complement(3342..3360)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3343..3360)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3380..3397)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3386..3610
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3656..4084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3743..3762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3953..3974
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4093..4113)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4098..4427
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4278..4413
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4340..4359
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4494..5288
                     /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(4548..4567)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5158..5177
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5462..5583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5499..5518)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5553..5572
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5632..5648)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5632..5648)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5645..5667)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5656..5672
                     /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(5680..5686,5687..5704,5705..5710))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5725..5746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5863..5880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6034..6622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6114..6133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6793..7584,7585..7653))
                     /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     7416..7435
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7654..7758)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7833..7852)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
       61 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
      121 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
      181 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      241 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      301 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      361 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      421 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      481 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      541 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      601 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      661 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      721 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
      781 cttatcgaaa ttaatacgac tcactatagg gagacccaag cttcgccacc atggtcttca
      841 cactcgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg gaccaagtcc
      901 ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta actccgatcc
      961 aaaggattgt cctgagcggt ctcgagggcg gtagcggtgg cagcgtgagc gtgatcaagc
     1021 ccgagatgaa gaaggtggag gacgccgtgg cccactccac tctcgagggc gtgagcgtga
     1081 tcaagcccga gatgaagatg aggtactaca tggacggctc cgtcaatggg catgagttca
     1141 caatcgaggg tgagggcaca ggcagacctt acgagggaca tcaggagatg acactgcgcg
     1201 tcacaatggc cgagggcggg ccaatgcctt tcgccttcga cctggtgtcc cacgtgttct
     1261 gttacggcca cagagttttt actaaatatc cagaagagat cccagactat ttcaagcagg
     1321 cctttcctga gggcctgtcc tgggagaggt ccctggagtt cgaggacggc ggctccgcct
     1381 ccgtgagcgc ccacatcagc ctgaggggca acaccttcta ccacaagtcc aagttcaccg
     1441 gcgtgaactt ccccgccgac ggccccatca tgcagaacca gagcgtggac tgggagccct
     1501 ccaccgagaa gatcaccgcc agcgacggcg tgctgaaggg cgacgtgacc atgtacctga
     1561 agctggaggg cggcggcaac cacaagtgcc agatgaagac cacctacaag gccgccaagg
     1621 agatcctgga gatgcccggc gaccactaca tcggccacag gctggtgagg aagaccgagg
     1681 gcaacatcac cgagcaggtg gaggacgccg tggcccactc cgagctcgaa aatgggctga
     1741 agatcgacat ccatgtcatc atcccgtatg aaggtctgag cggcgaccaa atgggccaga
     1801 tcgaaaaaat ttttaaggtg gtgtaccctg tggatgatca tcactttaag gtgatcctgc
     1861 actatggcac actggtaatc gacggggtta cgccgaacat gatcgactat ttcggacggc
     1921 cgtatgaagg catcgccgtg ttcgacggca aaaagatcac tgtaacaggg accctgtgga
     1981 acggcaacaa aattatcgac gagcgcctga tcaaccccga cggctccctg ctgttccgag
     2041 taaccatcaa cggagtgacc ggctggcggc tgtgcgaacg cattctggcg ggtggatcag
     2101 gcggtaccgg cagcggtggc ggtggcagcg gtggcggtgg cagcggtgga tccatggatg
     2161 atgatatcgc cgcgctcgtc gtcgacaacg gctccggcat gtgcaaggcc ggcttcgcgg
     2221 gcgacgatgc cccccgggcc gtcttcccct ccatcgtggg gcgccccagg caccagggcg
     2281 tgatggtggg catgggtcag aaggattcct atgtgggcga cgaggcccag agcaagagag
     2341 gcatcctcac cctgaagtac cccatcgagc acggcatcgt caccaactgg gacgacatgg
     2401 agaaaatctg gcaccacacc ttctacaatg agctgcgtgt ggctcccgag gagcaccccg
     2461 tgctgctgac cgaggccccc ctgaacccca aggccaaccg cgagaagatg acccagatca
     2521 tgtttgagac cttcaacacc ccagccatgt acgttgctat ccaggctgtg ctatccctgt
     2581 acgcctctgg ccgtaccact ggcatcgtga tggactccgg tgacggggtc acccacactg
     2641 tgcccatcta cgaggggtat gccctccccc atgccatcct gcgtctggac ctggctggcc
     2701 gggacctgac tgactacctc atgaagatcc tcaccgagcg cggctacagc ttcaccacca
     2761 cggccgagcg ggaaatcgtg cgtgacatta aggagaagct gtgctacgtc gccctggact
     2821 tcgagcaaga gatggccacg gctgcttcca gctcctccct ggagaagagc tacgagctgc
     2881 ctgacggcca ggtcatcacc attggcaatg agcggttccg ctgccctgag gcactcttcc
     2941 agccttcctt cctgggcatg gagtcctgtg gcatccacga aactaccttc aactccatca
     3001 tgaagtgtga cgtggacatc cgcaaagacc tgtacgccaa cacagtgctg tctggcggca
     3061 ccaccatgta ccctggcatt gccgacagga tgcagaagga gatcactgcc ctggcaccca
     3121 gcacaatgaa gatcaagatc attgctcctc ctgagcgcaa gtactccgtg tggatcggcg
     3181 gctccatcct ggcctcgctg tccaccttcc agcagatgtg gatcagcaag caggagtatg
     3241 acgagtccgg cccctccatc gtccaccgca aatgcttcta ggaattctgc agatatccat
     3301 cacactggcg gccgctcgag catgcatcta gagggcccta ttctatagtg tcacctaaat
     3361 gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3421 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3481 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3541 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3601 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3661 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3721 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3781 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3841 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3901 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3961 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4021 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4081 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4141 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4201 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4261 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4321 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4381 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4441 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4501 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4561 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4621 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4681 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4741 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4801 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4861 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4921 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4981 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5041 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5101 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5161 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5221 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5281 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5341 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5401 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5461 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5521 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5581 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5641 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5701 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5761 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5821 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5881 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5941 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6001 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6061 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6121 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6181 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6241 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6301 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6361 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6421 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6481 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6541 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6601 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6661 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6721 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6781 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6841 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6901 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6961 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7021 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7081 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7141 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7201 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7261 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7321 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7381 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7441 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7501 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7561 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7621 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7681 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7741 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7801 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
//
