LOCUS       sequence-233994-        5526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nucleus, Excitation: 551, Emission: 565.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5526)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 5526)
  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"
COMMENT     Sequence Label: mKO2-PCNA-19-SV40NLS-4
FEATURES             Location/Qualifiers
     source          1..5526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        110..413
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        414..617
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     568..588
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      644..653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             656..676
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             680..700
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             704..724
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      731..740
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             737..1390
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     polyA_signal    2363..2484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2400..2419)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2454..2473
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2491..2946)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2628..2649)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2840..2859
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2973..3077
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3079..3436
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3079..3270
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3102..3122)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3287..3422
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3471..4265
                     /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             3471..4265
                     /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(3525..3544)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4135..4154
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4453..4472)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4497..4544
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4873..5461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5362..5381
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
       61 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      121 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      181 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      241 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      301 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      361 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      421 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      481 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      541 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      601 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagccacca tggatccaaa
      661 aaagaagaga aaggtagatc caaaaaagaa gagaaaggta gatccaaaaa agaagagaaa
      721 ggtaccggtc gccaccatgg tgagtgtgat taaaccagag atgaagatga ggtactacat
      781 ggacggctcc gtcaatgggc atgagttcac aattgaaggt gaaggcacag gcagacctta
      841 cgagggacat caagagatga cactacgcgt cacaatggcc gagggcgggc caatgccttt
      901 cgcgtttgac ttagtgtcac acgtgttctg ttacggccac agagtattta ctaaatatcc
      961 agaagagata ccagactatt tcaaacaagc atttcctgaa ggcctgtcat gggaaaggtc
     1021 gttggagttc gaagatggtg ggtccgcttc agtcagtgcg catataagcc ttagaggaaa
     1081 caccttctac cacaaatcca aatttactgg ggttaacttt cctgccgatg gtcctatcat
     1141 gcaaaaccaa agtgttgatt gggagccatc aaccgagaaa attactgcca gcgacggagt
     1201 tctgaagggt gatgttacga tgtacctaaa acttgaagga ggcggcaatc acaaatgcca
     1261 aatgaagact acttacaagg cggcaaaaga gattcttgaa atgccaggag accattacat
     1321 cggccatcgc ctcgtcagga aaaccgaagg caacattact gagcaggtag aagatgcagt
     1381 agctcatagc tccggagaag ggcaagggca agggcaaggg ccgggccgcg gctacgcgta
     1441 tcgatccatg ttcgaggcgc gcctggtcca gggctccatc ctcaagaagg tgttggaggc
     1501 actcaaggac ctcatcaacg aggcctgctg ggatattagc tccagcggtg taaacctgca
     1561 gagcatggac tcgtcccacg tctctttggt gcagctcacc ctgcggtctg agggcttcga
     1621 cacctaccgc tgcgaccgca acctggccat gggcgtgaac ctcaccagta tgtccaaaat
     1681 actaaaatgc gccggcaatg aagatatcat tacactaagg gccgaagata acgcggatac
     1741 cttggcgcta gtatttgaag caccaaacca ggagaaagtt tcagactatg aaatgaagtt
     1801 gatggattta gatgttgaac aacttggaat tccagaacag gagtacagct gtgtagtaaa
     1861 gatgccttct ggtgaatttg cacgtatatg ccgagatctc agccatattg gagatgctgt
     1921 tgtaatttcc tgtgcaaaag acggagtgaa attttctgca agtggagaac ttggaaatgg
     1981 aaacattaaa ttgtcacaga caagtaatgt cgataaagag gaggaagctg ttaccataga
     2041 gatgaatgaa ccagttcaac taacttttgc actgaggtac ctgaacttct ttacaaaagc
     2101 cactccactc tcttcaacgg tgacactcag tatgtctgca gatgtacccc ttgttgtaga
     2161 gtataaaatt gcggatatgg gacacttaaa atactacttg gctcccaaga tcgaggatga
     2221 agaaggatct tagggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     2281 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2341 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2401 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2461 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     2521 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2581 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2641 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2701 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2761 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2821 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2881 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2941 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3001 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3061 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3121 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3181 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3241 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3301 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3361 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3421 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3481 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3541 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3601 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3661 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3721 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3781 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3841 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3901 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3961 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4021 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4081 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4141 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4201 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4261 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4321 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4381 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4441 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4501 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4561 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4621 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4681 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     4741 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4801 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4861 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4921 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4981 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5041 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5101 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5161 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5221 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5281 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5341 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5401 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5461 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5521 gttctt
//
