LOCUS       sequence-115521-        5603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ONL-PTS1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5603)
  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 5603)
  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_PTS1
FEATURES             Location/Qualifiers
     source          1..5603
                     /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"
     polyA_signal    2391..2512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2428..2447)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2482..2501
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2519..2974)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2656..2677)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2868..2887
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3001..3105
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3107..3464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3107..3298
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3130..3150)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3315..3450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3499..4293
                     /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             3499..4293
                     /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(3553..3572)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4163..4182
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4481..4500)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4525..4572
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4901..5489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5390..5409
                     /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 ccgggatatc caagctgtga gtcgacggta ccgcgggccc gggatccacc ggatctagat
     2281 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2341 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2401 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2461 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     2521 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2581 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2641 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2701 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2761 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2821 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2881 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2941 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3001 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3061 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3121 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3181 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3241 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3301 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3361 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3421 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     3481 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3541 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3601 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3661 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     3721 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3781 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3841 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3901 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3961 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4021 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4081 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4141 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4201 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4261 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4321 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4381 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4441 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     4501 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     4561 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4621 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4681 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     4741 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4801 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4861 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4921 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4981 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5041 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5101 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5161 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5221 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5281 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5341 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5401 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5461 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5521 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5581 ataaccgtat taccgccatg cat
//
