LOCUS       sequence-397396-        5583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKO-tagged human importin-alpha in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5583)
  AUTHORS   Nakagawa C, Nishimura S, Senda-Murata K, Sugimoto K
  TITLE     A rapid and simple method of evaluating the dimeric tendency of 
            fluorescent proteins in living cells using a truncated protein of 
            importin alpha as fusion tag.
  JOURNAL   Biosci Biotechnol Biochem. 2012;76(2):388-90. doi: 
            10.1271/bbb.110677. Epub 2012 Feb 7.
  PUBMED    22313767
REFERENCE   2  (bases 1 to 5583)
  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"; journalName: "Biosci 
            Biotechnol Biochem."; date: "2012"; volume: "76(2):388-90. doi"; 
            pages: " 10.1271/bbb.110677. Epub 2012 Feb 7"
COMMENT     Sequence Label: pmKO-imp alpha
FEATURES             Location/Qualifiers
     source          1..5583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             189..983
                     /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             189..983
                     /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(243..262)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     853..872
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1171..1190)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1215..1262
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1591..2179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2080..2099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2354..2657
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2658..2861
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2812..2832
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2893..2902
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2899..3552
                     /codon_start=1
                     /product="monomeric Kusabira-Orange fluorescent protein 
                     (Karasawa et al., 2004)"
                     /label=mKO
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     polyA_signal    4665..4786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4702..4721)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4756..4775
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4793..5248)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4930..4951)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5142..5161
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5275..5379
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5381..5583,1..155)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5381..5572
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5404..5424)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
       61 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
      121 gaggaggctt ttttggaggc ctaggctttt gcaaagatca tcaagagaca ggatgaggat
      181 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
      241 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
      301 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
      361 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
      421 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
      481 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
      541 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
      601 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
      661 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
      721 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
      781 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
      841 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
      901 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
      961 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     1021 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     1081 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     1141 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     1201 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     1261 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     1321 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     1381 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     1441 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1501 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1561 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1621 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1681 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1741 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1801 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1861 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1921 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1981 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2041 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2101 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2161 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2221 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2281 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     2341 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2401 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2461 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2521 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2581 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2641 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2701 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2761 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2821 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     2881 tccgctagca ttgccaccat ggtgagtgtg attaaaccag agatgaagat gaggtactac
     2941 atggacggct ccgtcaatgg gcatgagttc acaattgaag gtgaaggcac aggcagacct
     3001 tacgagggac atcaagagat gacactacgc gtcacaatgg ccaagggcgg gccaatgcct
     3061 ttcgcgtttg acttagtgtc acacgtgttc tgttacggcc acagaccttt tactaaatat
     3121 ccagaagaga taccagacta tttcaaacaa gcatttcctg aaggcctgtc atgggaaagg
     3181 tcgttggagt tcgaagatgg tgggtccgct tcagtcagtg cgcatataag ccttagagga
     3241 aacaccttct accacaaatc caaatttact ggggttaact ttcctgccga tggtcctatc
     3301 atgcaaaacc aaagtgttga ttgggagcca tcaaccgaga aaattactgc cagcgacgga
     3361 gttctgaagg gtgatgttac gatgtaccta aaacttgaag gaggcggcaa tcacaaatgc
     3421 caattcaaga ctacttacaa ggcggcaaaa aagattctta aaatgccagg aagccattac
     3481 atcagccatc gcctcgtcag gaaaaccgaa ggcaacatta ctgagctggt agaagatgca
     3541 gtagctcatt ccggatctcg agctcaagct agcttcgaat tcggcacgag gcagattctt
     3601 cctaccttag ttcggctcct gcatcatgat gatccagaag tgttagcaga tacctgctgg
     3661 gctatttcct accttactga tggtccaaat gaacgaattg gcatggtggt gaaaacagga
     3721 gttgtgcccc aacttgtgaa gcttctagga gcttctgaat tgccaattgt gactcctgcc
     3781 ctaagagcca tagggaatat tgtcactggt acagatgaac agactcaggt tgtgattgat
     3841 gcaggagcac tcgccgtctt tcccagcctg ctcaccaacc ccaaaactaa cattcagaag
     3901 gaagctacgt ggacaatgtc aaacatcaca gccggccgcc aggaccagat acagcaagtt
     3961 gtgaatcatg gattagtccc attccttgtc agtgttctct ctaaggcaga ttttaagaca
     4021 caaaaggaag ctgtgtgggc cgtgaccaac tataccagtg gtggaacagt tgaacagatt
     4081 gtgtaccttg ttcactgtgg cataatagaa ccgttgatga acctcttaac tgcaaaagat
     4141 accaagatta ttctggttat cctggatgcc atttcaaata tctttcaggc tgctgagaaa
     4201 ctaggtgaaa ctgagaaact tagtataatg attgaagaat gtggaggctt agacaaaatt
     4261 gaagctctac aaaaccatga aaatgagtct gtgtataagg cttcgttaag cttaattgag
     4321 aagtatttct ctgtagagga agaggaagat caaaacgttg taccagaaac tacctctgaa
     4381 ggctacactt tccaagttca ggatggggct cctgggacct ttaactttta gatcatgtag
     4441 ctgagacata aatttgttgt gtactaaaaa aaaaaaaaaa aaactcgacg gtaccggaac
     4501 tgcagcagag aattcgggaa actcgagaac aaagcttgaa taagcggccg cgactctaga
     4561 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4621 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     4681 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4741 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     4801 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4861 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4921 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4981 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5041 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5101 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     5161 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     5221 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5281 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5341 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     5401 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5461 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5521 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5581 ctc
//
