LOCUS       sequence-115495-        6197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ONL-RhoB in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6197)
  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 6197)
  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_RhoB
FEATURES             Location/Qualifiers
     source          1..6197
                     /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"
     CDS             2223..2252
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2985..3106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3022..3041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3076..3095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3113..3568)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3250..3271)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3462..3481
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3595..3699
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3701..4058
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3701..3892
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3724..3744)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3909..4044
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4093..4887
                     /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             4093..4887
                     /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(4147..4166)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4757..4776
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5075..5094)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5119..5166
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5495..6083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5984..6003
                     /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 ccgagcagaa gctgatctcc gaggaggacc tgaaccggaa gaaactggtg gttgttggtg
     2281 atggagcctg tggaaagaca tgcttgctca tagtcttcag caaggacgag ttcccagagg
     2341 tgtatgtgcc cacagtgttt gagaactatg tggcagatat cgaggtggat ggaaagcagg
     2401 tagagttggc tttgtgggac acagctggcc aggaggacta cgaccgcctg cggccgctct
     2461 cctacccgga caccgacgtc attctcatgt gcttctcggt ggacagcccg gactcgctgg
     2521 agaacatccc cgagaagtgg gtccccgagg tgaagcactt ctgtcccaat gtgcccatca
     2581 tcctggtggc caacaaaaaa gacctgcgca gcgacgagca tgtccgcaca gagctggccc
     2641 gcatgaagca ggaacccgtt cgcacggatg acggccgcgc catggccgtg cgcatccaag
     2701 cctacgacta cctcgagtgc tctgccaaga ccaaggaagg cgtgcgcgag gtcttcgaga
     2761 cggccacgcg cgccgcgctg cagaagcgct acggctccca gaacggctgc atcaactgct
     2821 gcaaggtgct atgagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     2881 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2941 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3001 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3061 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     3121 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3181 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3241 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3301 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3361 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3421 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3481 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3541 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3601 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3661 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3721 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3781 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3841 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3901 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3961 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4021 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4081 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4141 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4201 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4261 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4321 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4381 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4441 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4501 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4561 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4621 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4681 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4741 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4801 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4861 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4921 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4981 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5041 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5101 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5161 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5221 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5281 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5341 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5401 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5461 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5521 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5581 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5641 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5701 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5761 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5821 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5881 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5941 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6001 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6061 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6121 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6181 gtattaccgc catgcat
//
