LOCUS       sequence-177777-        6797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Orange color variant of bright luminescent protein, targeted to the 
            plasma membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6797)
  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 6797)
  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: Lyn-OeNL_pcDNA3
FEATURES             Location/Qualifiers
     source          1..6797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        132..511
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        512..715
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     666..686
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     760..779
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        760..778
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      792..801
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             882..899
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             903..935
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(939..947,948..962)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1209..1859
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             1209..1859
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     promoter        complement(2247..2265)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2248..2265)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2285..2302)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2291..2515
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2561..2989
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2648..2667)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2858..2879
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2998..3018)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3003..3332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3183..3318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3245..3264
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3399..4193
                     /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(3453..3472)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4063..4082
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4367..4488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4404..4423)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4458..4477
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4537..4553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4537..4553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4550..4572)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4561..4577
                     /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(4585..4591,4592..4609,4610..4615))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4630..4651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4768..4785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4939..5527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5019..5038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5698..6489,6490..6558))
                     /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     6321..6340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6559..6663)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6738..6757)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
       61 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
      121 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      181 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      241 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      301 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      361 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      421 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      481 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      541 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      601 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      661 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
      721 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
      781 gacccaagct tgccaccatg ggctgcatca agagcaagcg caaggacaac ctgaacgacg
      841 acggcgtgga catgaagacc ggcggccgca tgcggggttc tcatcatcat catcatcatg
      901 gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata
      961 aggatccgat ggtcttcaca ctcgaagatt tcgttgggga ctggcgacag acagccggct
     1021 acaacctgga ccaagtcctt gaacagggag gtgtgtccag tttgtttcag aatctcgggg
     1081 tgtccgtaac tccgatccaa aggattgtcc tgagcggtct cgagggcggt agcggtggca
     1141 gcgtgagcgt gatcaagccc gagatgaaga aggtggagga cgccgtggcc cactccactc
     1201 tcgagggcgt gagcgtgatc aagcccgaga tgaagatgag gtactacatg gacggctccg
     1261 tcaatgggca tgagttcaca atcgagggtg agggcacagg cagaccttac gagggacatc
     1321 aggagatgac actgcgcgtc acaatggccg agggcgggcc aatgcctttc gccttcgacc
     1381 tggtgtccca cgtgttctgt tacggccaca gagtttttac taaatatcca gaagagatcc
     1441 cagactattt caagcaggcc tttcctgagg gcctgtcctg ggagaggtcc ctggagttcg
     1501 aggacggcgg ctccgcctcc gtgagcgccc acatcagcct gaggggcaac accttctacc
     1561 acaagtccaa gttcaccggc gtgaacttcc ccgccgacgg ccccatcatg cagaaccaga
     1621 gcgtggactg ggagccctcc accgagaaga tcaccgccag cgacggcgtg ctgaagggcg
     1681 acgtgaccat gtacctgaag ctggagggcg gcggcaacca caagtgccag atgaagacca
     1741 cctacaaggc cgccaaggag atcctggaga tgcccggcga ccactacatc ggccacaggc
     1801 tggtgaggaa gaccgagggc aacatcaccg agcaggtgga ggacgccgtg gcccactccg
     1861 agctcgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg
     1921 gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc
     1981 actttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg ccgaacatga
     2041 tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg
     2101 taacagggac cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg
     2161 gctccctgct gttccgagta accatcaacg gagtgaccgg ctggcggctg tgcgaacgca
     2221 ttctggcgta atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     2281 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2341 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2401 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2461 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2521 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2581 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2641 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2701 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2761 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2821 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2881 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2941 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3001 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3061 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3121 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3181 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3241 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3301 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3361 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3421 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3481 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3541 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3601 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3661 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3721 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3781 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3841 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3901 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3961 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4021 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4081 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4141 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4201 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4261 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4321 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4381 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4441 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4501 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4561 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4621 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4681 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4741 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4801 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4861 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4921 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4981 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5041 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5101 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5161 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5221 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5281 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5341 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5401 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5461 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5521 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5581 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5641 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5701 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5761 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5821 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5881 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5941 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6001 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6061 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6121 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6181 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6241 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6301 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6361 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6421 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6481 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6541 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6601 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6661 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     6721 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     6781 cctgcttgtg tgttgga
//
