LOCUS       sequence-236000-        5605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mOrange-tagged human importin-alpha in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5605)
  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 5605)
  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: mOrange-imp alpha
FEATURES             Location/Qualifiers
     source          1..5605
                     /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      598..607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             604..1311
                     /codon_start=1
                     /product="orange monomeric derivative of DsRed fluorescent 
                     protein (Shanet et al., 2004)"
                     /label=mOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1017..1036
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2393..2514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2430..2449)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2484..2503
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2521..2976)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2658..2679)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2870..2889
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3003..3107
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3109..3466
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3109..3300
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3132..3152)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3317..3452
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3501..4295
                     /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             3501..4295
                     /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(3555..3574)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4165..4184
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4483..4502)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4527..4574
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4903..5491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5392..5411
                     /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 gctagccgcc
      601 accatggtga gcaagggcga ggagaataac atggccatca tcaaggagtt catgcgcttc
      661 aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
      721 ggccgcccct acgagggctt tcagaccgct aagctgaagg tgaccaaggg tggccccctg
      781 cccttcgcct gggacatcct gtcccctcag ttcacctacg gctccaaggc ctacgtgaag
      841 caccccgccg acatccccga ctacttcaag ctgtccttcc ccgagggctt caagtgggag
      901 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
      961 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1021 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     1081 ggcgccctga agggcgagat caagatgagg ctgaagctga aggacggcgg ccactacacc
     1141 tccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacatc
     1201 gtcggcatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1261 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gagatctcga
     1321 gctcaagcta gcttcgaatt cggcacgagg cagattcttc ctaccttagt tcggctcctg
     1381 catcatgatg atccagaagt gttagcagat acctgctggg ctatttccta ccttactgat
     1441 ggtccaaatg aacgaattgg catggtggtg aaaacaggag ttgtgcccca acttgtgaag
     1501 cttctaggag cttctgaatt gccaattgtg actcctgccc taagagccat agggaatatt
     1561 gtcactggta cagatgaaca gactcaggtt gtgattgatg caggagcact cgccgtcttt
     1621 cccagcctgc tcaccaaccc caaaactaac attcagaagg aagctacgtg gacaatgtca
     1681 aacatcacag ccggccgcca ggaccagata cagcaagttg tgaatcatgg attagtccca
     1741 ttccttgtca gtgttctctc taaggcagat tttaagacac aaaaggaagc tgtgtgggcc
     1801 gtgaccaact ataccagtgg tggaacagtt gaacagattg tgtaccttgt tcactgtggc
     1861 ataatagaac cgttgatgaa cctcttaact gcaaaagata ccaagattat tctggttatc
     1921 ctggatgcca tttcaaatat ctttcaggct gctgagaaac taggtgaaac tgagaaactt
     1981 agtataatga ttgaagaatg tggaggctta gacaaaattg aagctctaca aaaccatgaa
     2041 aatgagtctg tgtataaggc ttcgttaagc ttaattgaga agtatttctc tgtagaggaa
     2101 gaggaagatc aaaacgttgt accagaaact acctctgaag gctacacttt ccaagttcag
     2161 gatggggctc ctgggacctt taacttttag atcatgtagc tgagacataa atttgttgtg
     2221 tactaaaaaa aaaaaaaaaa aactcgacgg taccgcgggc ccgggatcca ccggatctag
     2281 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2341 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2401 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2461 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     2521 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2581 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2641 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2701 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2761 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2821 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2881 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     2941 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3001 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3061 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3121 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3181 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3241 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3301 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3361 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3421 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3481 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3541 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3601 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3661 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     3721 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3781 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3841 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3901 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3961 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4021 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4081 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4141 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4201 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4261 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4321 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4381 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4441 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4501 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4561 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4621 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4681 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     4741 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4801 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4861 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4921 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4981 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5041 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5101 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5161 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5221 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5281 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5341 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5401 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5461 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5521 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5581 ggataaccgt attaccgcca tgcat
//
