LOCUS       sequence-177724-        8047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Orange color variant of bright luminescent protein, fused to 
            b-tubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8047)
  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 8047)
  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: tubulin-OeNL_pcDNA3
FEATURES             Location/Qualifiers
     source          1..8047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        133..512
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        513..716
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     667..687
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     761..780
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        761..779
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2441..3091
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             2441..3091
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     promoter        complement(3498..3516)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3499..3516)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3536..3553)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3542..3766
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3812..4240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3899..3918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4109..4130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4249..4269)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4254..4583
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4434..4569
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4496..4515
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4650..5444
                     /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(4704..4723)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5314..5333
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5618..5739
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5655..5674)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5709..5728
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5788..5804)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5788..5804)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5801..5823)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5812..5828
                     /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(5836..5842,5843..5860,5861..5866))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5881..5902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6019..6036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6190..6778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6270..6289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6949..7740,7741..7809))
                     /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     7572..7591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7810..7914)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7989..8008)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
       61 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
      121 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      181 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      241 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      301 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      361 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      421 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      481 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      541 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      601 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      661 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct
      721 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
      781 agacccaagc ttcgccacca tgagggaaat cgtgcacatc caggccggac agtgtggcaa
      841 ccagatcggt gctaagttct gggaggtgat aagcgatgaa catggcatcg accccaccgg
      901 tacctaccac ggtgacagcg acctgcagct ggaccgaatc tctgtgtact ataatgaagc
      961 cacaggtggc aagtatgtcc ctcgagctat cttggtggat ctagaacctg ggactatgga
     1021 ctccgttcgc tcaggtcctt ttggccagat cttcagacca gacaacttcg ttttcggtca
     1081 gtctggggca ggcaacaact gggctaaagg ccactacaca gagggagctg agttggttga
     1141 ctctgtcttg gatgtggtgc ggaaggaggc ggagagctgt gattgcctgc aaggctttca
     1201 gctgacccac tcactgggtg gaggcactgg ctctggcatg ggcaccctgc tcatcagcaa
     1261 gatccgggaa gaatatcctg accgtatcat gaataccttc agtgtggtgc cctcgcccaa
     1321 agtgtctgat accgtggtcg agccctacaa tgccaccctg tctgtccatc agttggttga
     1381 gaacacggat gagacctact gcatcgacaa cgaggccctc tacgacatct gcttccgtac
     1441 cctcaagctc accacgccaa cctacggaga cctgaaccat ctcgtctcgg ccaccatgag
     1501 cggcgtcacc acctgcctcc gtttcccggg ccagcttaat gctgaccttc gaaagctggc
     1561 tgtcaacatg gtgccattcc cacgtctcca cttcttcatg cctggctttg cccctctcac
     1621 cagccgtgga agccagcagt accgggccct cactgtgcct gaacttaccc agcaggtctt
     1681 cgatgccaag aacatgatgg ccgcctgcga cccgcgccac ggccggtacc tcacagttgc
     1741 cgccgtcttc cgtggacgga tgtccatgaa ggaggtggat gagcagatgc tcaacgtgca
     1801 gaacaagaat agcagctact tcgtggaatg gatccccaac aatgtcaaga cagctgtctg
     1861 tgacatccca ccgcgtggcc tcaagatggc agtcaccttc attggaaaca gcacagccat
     1921 ccaggagctc ttcaagcgca tctctgagca gtttacggct atgttccgcc ggaaggcttt
     1981 cctccactgg tatacgggtg agggcatgga cgagatggag ttcaccgaag ctgagagcaa
     2041 catgaacgac ctcgtctctg agtaccagca gtaccaggat gccaccgcgg aagaggaaga
     2101 ggatttcgga gaggaggcag aagaggaggc ccagtcgacg gtaccgcggg cccgggatcc
     2161 aggatctggc ggaggaagtg ggggaggatc tggagaattc atggtcttca cactcgaaga
     2221 tttcgttggg gactggcgac agacagccgg ctacaacctg gaccaagtcc ttgaacaggg
     2281 aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta actccgatcc aaaggattgt
     2341 cctgagcggt ctcgagggcg gtagcggtgg cagcgtgagc gtgatcaagc ccgagatgaa
     2401 gaaggtggag gacgccgtgg cccactccac tctcgagggc gtgagcgtga tcaagcccga
     2461 gatgaagatg aggtactaca tggacggctc cgtcaatggg catgagttca caatcgaggg
     2521 tgagggcaca ggcagacctt acgagggaca tcaggagatg acactgcgcg tcacaatggc
     2581 cgagggcggg ccaatgcctt tcgccttcga cctggtgtcc cacgtgttct gttacggcca
     2641 cagagttttt actaaatatc cagaagagat cccagactat ttcaagcagg cctttcctga
     2701 gggcctgtcc tgggagaggt ccctggagtt cgaggacggc ggctccgcct ccgtgagcgc
     2761 ccacatcagc ctgaggggca acaccttcta ccacaagtcc aagttcaccg gcgtgaactt
     2821 ccccgccgac ggccccatca tgcagaacca gagcgtggac tgggagccct ccaccgagaa
     2881 gatcaccgcc agcgacggcg tgctgaaggg cgacgtgacc atgtacctga agctggaggg
     2941 cggcggcaac cacaagtgcc agatgaagac cacctacaag gccgccaagg agatcctgga
     3001 gatgcccggc gaccactaca tcggccacag gctggtgagg aagaccgagg gcaacatcac
     3061 cgagcaggtg gaggacgccg tggcccactc cgagctcgaa aatgggctga agatcgacat
     3121 ccatgtcatc atcccgtatg aaggtctgag cggcgaccaa atgggccaga tcgaaaaaat
     3181 ttttaaggtg gtgtaccctg tggatgatca tcactttaag gtgatcctgc actatggcac
     3241 actggtaatc gacggggtta cgccgaacat gatcgactat ttcggacggc cgtatgaagg
     3301 catcgccgtg ttcgacggca aaaagatcac tgtaacaggg accctgtgga acggcaacaa
     3361 aattatcgac gagcgcctga tcaaccccga cggctccctg ctgttccgag taaccatcaa
     3421 cggagtgacc ggctggcggc tgtgcgaacg cattctggcg taagcggccg ctcgagcatg
     3481 catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg
     3541 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3601 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3661 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3721 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3781 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3841 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3901 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3961 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4021 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4081 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4141 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4201 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4261 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4321 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4381 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4441 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4501 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4561 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4621 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4681 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4741 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4801 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4861 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4921 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4981 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5041 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5101 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5161 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5221 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5281 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5341 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5401 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5461 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5521 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5581 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5641 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5701 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5761 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5821 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5881 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5941 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6001 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6061 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7921 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     7981 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     8041 gtgttgg
//
