LOCUS       sequence-162741-        4669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: N1 Cloning Vector, Excitation: 551, Emission: 565.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4669)
  AUTHORS   Sakaue-Sawano A, Kurokawa H, Morimura T, Hanyu A, Hama H, Osawa H, 
            Kashiwagi S, Fukami K, Miyata T, Miyoshi H, Imamura T, Ogawa M, 
            Masai H, Miyawaki A
  TITLE     Visualizing spatiotemporal dynamics of multicellular cell-cycle 
            progression.
  JOURNAL   Cell. 2008 Feb 8;132(3):487-98. doi: 10.1016/j.cell.2007.12.033.
  PUBMED    18267078
REFERENCE   2  (bases 1 to 4669)
  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: 
            "10.1016/j.cell.2007.12"; journalName: "Cell"; date: "2008-02-8- 8";
            volume: "132"; issue: "3"; pages: "487-98"
COMMENT     Sequence Label: mKO2-N1
FEATURES             Location/Qualifiers
     source          1..4669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    546..626
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             634..1290
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     polyA_signal    1412..1533
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1449..1468)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1503..1522
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1540..1995)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1677..1698)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1889..1908
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2022..2126
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2128..2485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2128..2319
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2151..2171)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2336..2471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2520..3314
                     /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             2520..3314
                     /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(2574..2593)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3184..3203
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3502..3521)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3546..3593
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3922..4510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4411..4430
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgacg
      601 gtaccgcggg cccgggatcc accggtcgcc accatggtga gtgtgattaa accagagatg
      661 aagatgaggt actacatgga cggctccgtc aatgggcatg agttcacaat tgaaggtgaa
      721 ggcacaggca gaccttacga gggacatcaa gagatgacac tacgcgtcac aatggccgag
      781 ggcgggccaa tgcctttcgc gtttgactta gtgtcacacg tgttctgtta cggccacaga
      841 gtatttacta aatatccaga agagatacca gactatttca aacaagcatt tcctgaaggc
      901 ctgtcatggg aaaggtcgtt ggagttcgaa gatggtgggt ccgcttcagt cagtgcgcat
      961 ataagcctta gaggaaacac cttctaccac aaatccaaat ttactggggt taactttcct
     1021 gccgatggtc ctatcatgca aaaccaaagt gttgattggg agccatcaac cgagaaaatt
     1081 actgccagcg acggagttct gaagggtgat gttacgatgt acctaaaact tgaaggaggc
     1141 ggcaatcaca aatgccaaat gaagactact tacaaggcgg caaaagagat tcttgaaatg
     1201 ccaggagacc attacatcgg ccatcgcctc gtcaggaaaa ccgaaggcaa cattactgag
     1261 caggtagaag atgcagtagc tcatagctaa gcggccgcga ctctagatca taatcagcca
     1321 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     1381 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     1441 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1501 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     1561 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     1621 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     1681 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     1741 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     1801 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     1861 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     1921 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     1981 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2041 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2101 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     2161 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2221 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2281 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2341 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2401 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     2461 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     2521 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2581 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     2641 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     2701 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     2761 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     2821 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     2881 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     2941 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3001 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     3061 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3121 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3181 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3241 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3301 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     3361 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3421 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3481 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     3541 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     3601 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     3661 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     3721 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     3781 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3841 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3901 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3961 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4021 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4081 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4141 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4201 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4261 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4321 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4381 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4441 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4501 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4561 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     4621 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccc
//
