LOCUS       sequence-225111-        4656 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Kusabira- Orange-CENP-A expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4656)
  AUTHORS   Sugimoto K, Senda-Murata K, Oka S
  TITLE     Construction of three quadruple-fluorescent MDA435 cell lines that 
            enable monitoring of the whole chromosome segregation process in the
            living state.
  JOURNAL   Mutat Res. 2008 Nov 17;657(1):56-62. doi: 
            10.1016/j.mrgentox.2008.08.005. Epub 2008 Aug 19.
  PUBMED    18778791
REFERENCE   2  (bases 1 to 4656)
  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.mrgentox.2008.08.005"; journalName: "Mutat Res"; date: 
            "2008-11-17- 17"; volume: "657"; issue: "1"; pages: "56-62"
COMMENT     Sequence Label: pmKO-AF8
FEATURES             Location/Qualifiers
     source          1..4656
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..654
                     /codon_start=1
                     /product="monomeric Kusabira-Orange fluorescent protein 
                     (Karasawa et al., 2004)"
                     /label=mKO
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     polyA_signal    839..960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(876..895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     930..949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(967..1422)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1104..1125)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1316..1335
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1449..1553
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1555..1912
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1555..1746
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1578..1598)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1763..1898
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1947..2741
                     /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             1947..2741
                     /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(2001..2020)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2611..2630
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2929..2948)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2973..3020
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3349..3937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3838..3857
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4112..4415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4416..4619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4570..4590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atggtgagtg tgattaaacc agagatgaag atgaggtact acatggacgg ctccgtcaat
       61 gggcatgagt tcacaattga aggtgaaggc acaggcagac cttacgaggg acatcaagag
      121 atgacactac gcgtcacaat ggccaagggc gggccaatgc ctttcgcgtt tgacttagtg
      181 tcacacgtgt tctgttacgg ccacagacct tttactaaat atccagaaga gataccagac
      241 tatttcaaac aagcatttcc tgaaggcctg tcatgggaaa ggtcgttgga gttcgaagat
      301 ggtgggtccg cttcagtcag tgcgcatata agccttagag gaaacacctt ctaccacaaa
      361 tccaaattta ctggggttaa ctttcctgcc gatggtccta tcatgcaaaa ccaaagtgtt
      421 gattgggagc catcaaccga gaaaattact gccagcgacg gagttctgaa gggtgatgtt
      481 acgatgtacc taaaacttga aggaggcggc aatcacaaat gccaattcaa gactacttac
      541 aaggcggcaa aaaagattct taaaatgcca ggaagccatt acatcagcca tcgcctcgtc
      601 aggaaaaccg aaggcaacat tactgagctg gtagaagatg cagtagctca ttccggatcc
      661 tcaggtaccg gaactgcagc agagaattcg ggaaactcga gaacaaagct tgaataagcg
      721 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
      781 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
      841 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
      901 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
      961 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     1021 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     1081 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     1141 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     1201 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     1261 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     1321 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     1381 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     1441 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1501 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     1561 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     1621 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     1681 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     1741 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     1801 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     1861 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     1921 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     1981 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2041 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2101 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     2161 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2221 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     2281 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     2341 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     2401 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     2461 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     2521 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     2581 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     2641 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     2701 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     2761 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     2821 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     2881 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     2941 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     3001 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     3061 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3121 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3181 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3241 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3301 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3361 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3421 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3481 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3541 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3601 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3661 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3721 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3781 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3841 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3901 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3961 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4021 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     4081 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4141 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4201 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4261 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4321 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4381 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     4441 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     4501 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     4561 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     4621 gtttagtgaa ccgtcagatc cgctagcatt gccacc
//
