LOCUS       sequence-391901-        7409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  visualization of nucleolus in living mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7409)
  AUTHORS   Fukada T, Shibata S, Ueda T, Sasaki K, Shimoida Y, Senda-Murata K, 
            Sugimoto K
  TITLE     Characterization of nucleolar localization and exclusion signals in 
            terminal deoxynucleotidyltransferase interacting factor 2/estrogen 
            receptor alpha-binding protein.
  JOURNAL   Biosci Biotechnol Biochem. 2019 Jul;83(7):1255-1262. doi: 
            10.1080/09168451.2019.1591265. Epub 2019 Mar 24.
  PUBMED    30907250
REFERENCE   2  (bases 1 to 7409)
  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: 
            "10.1080/09168451.2019.1591265"; journalName: "Biosci Biotechnol 
            Biochem"; date: "2019-07"; volume: "83"; issue: "7"; pages: 
            "1255-1262"
COMMENT     Sequence Label: pEGFP-TdIF2
FEATURES             Location/Qualifiers
     source          1..7409
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1366..1386
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1393..1629
                     /codon_start=1
                     /product="yeast transcriptional activator cretaed from E. 
                     coli genomic DNA fragments (Ma and Ptashne, 1987)"
                     /label=B42 transcriptional activator
                     /translation="INKDIEECNAIIEQFIDYLRTGQEMPMEMADQAINVVPGMTPKTI
                     LHAGPPIQPDWLKSNGFHEIEADVNDTSLLLSGD"
     CDS             1636..1662
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4197..4318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4234..4253)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4288..4307
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4325..4780)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4462..4483)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4674..4693
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4807..4911
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4913..5270
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4913..5104
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4936..4956)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5121..5256
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5305..6099
                     /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             5305..6099
                     /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(5359..5378)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5969..5988
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6287..6306)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6331..6378
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6707..7295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7196..7215
                     /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 gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt actcagatct cgagctcaag cttatgggtg ctcctccaaa aaagaagaga
     1381 aaggtagctg gtatcaataa agatatcgag gagtgcaatg ccatcattga gcagtttatc
     1441 gactacctgc gcaccggaca ggagatgccg atggaaatgg cggatcaggc gattaacgtg
     1501 gtgccgggca tgacgccgaa aaccattctt cacgccgggc cgccgatcca gcctgactgg
     1561 ctgaaatcga atggttttca tgaaattgaa gcggatgtta acgataccag cctcttgctg
     1621 agtggagatg cctcctaccc ttatgatgtg ccagattatg cctctcccga attcggcacg
     1681 agaaagatgg tggttaccag atctgcacgg gctaaggcca gcatccaagc cgcgtcggct
     1741 gaaagttccg ggcaaaagag ttttgctgct aatgggattc aagcgcatcc agaaagtagt
     1801 actggatctg atgcccgaac tactgctgaa tcacagacca ctgggaagca aagtttaatc
     1861 cctagaactc ctaaagctag aaagaggaag agcagaacta caggctcact accaaagggg
     1921 actgaaccat ctacggatgg agaaacctct gaggcagagt caaattattc tgtgtctgag
     1981 caccatgata ccattttaag ggtaactagg agaaggcaga tcttaattgc atgctcccca
     2041 gtgtccagtg ttaggaaaaa gccgaaagta actccaacaa aggagtctta cactgaagaa
     2101 atagtgtctg aagcagaatc tcatgtttca ggtatttcta gaattgtgct tcctacagaa
     2161 aaaactacag gagccagaag aagtaaggct aaatctctga cagatccaag ccaagaatct
     2221 catacagaag ctatatctga tgctgagaca tcaagctcag acatttcatt ctctggaatt
     2281 gcaactagaa gaaccaggag tatgcagagg aaattaaagg cacaaactga aaagaaagat
     2341 agtaagattg taccaggaaa tgagaaacag atcgtgggta cacctgtgaa ttcagaggat
     2401 tcagatacca gacaaacttc ccatttacaa gcaagatctc tttctgagat aaataagcca
     2461 aatttctata ataatgactt tgatgatgat ttctcccaca gaagttcaga aaatatatta
     2521 acagtgcacg aacaggccaa tgttgaatct cttaaagaaa caaaacagaa ttgtaaggat
     2581 ttggatgaag atgccaatgg aataacagat gaggggaaag aaattaatga gaaaagttct
     2641 cagctgaaga atctttctga acttcaggac actagccttc aacagttagt ttctcagaga
     2701 cattcaaccc cccaaaataa aaatgctgta tcagtgcact ctaatctgaa ctctgaggct
     2761 gtaatgaaat cattaactca aacatttgca actgtggaag taggcagatg gaataacaac
     2821 aaaaagagcc ccataaaagc aagtgacttg acaaagtttg gtgattgtgg tggtagtgat
     2881 gatgaagaag agtccacagt tataagtgtc agtgaagaca tgaacagtga agggaatgta
     2941 gattttgaat gtgataccaa actatacacg tctgcgccca acacatctca gggtaaagat
     3001 aattctgtct tactagttct cagcagtgat gaaagccaac agtctgaaaa cagtgagaat
     3061 gaagaggata ctttatgttt tgttgaaaat agtggccaaa gggagtcatt aagtggagac
     3121 acaggaagtc tgtcatgtga caatgcattg tttgtaattg acacaactcc tggaatgagt
     3181 gctgataaaa atttttactt ggaagaggaa gacaaggcaa gtgaggttgc cattgaggaa
     3241 gaaaaagaag aggaagagga tgaaaaaagt gaagaagatt catcagacca tgacgaaaat
     3301 gaagatgagt ttagtgatga agaagacttc ctaaatagca caaaggctaa acttctgaag
     3361 ttgacaagca gcagcataga ccctggtctg agtatcaagc agttgggtgg tttgtatatt
     3421 aattttaatg cagataaact acagtctaac aagagaaccc taacacagat caaggagaaa
     3481 aagaaaaatg agcttctgca gaaagccgtc attacacctg attttgaaaa aaaccactgt
     3541 gttccaccat atagtgaatc aaagtatcaa cttcagaaaa aacgcagaaa agaacgacaa
     3601 aaaacagcag gggatggctg gtttggtatg aaagctccag aaatgacaaa tgaactgaaa
     3661 aatgatctca aagcactgaa gatgagagcc agcatggacc cgaaaagatt ttacaagaaa
     3721 aatgatagag atggcttccc caagtacttc cagattggaa ccattgttga caatccagct
     3781 gatttctacc attcacgaat tcccaagaag caaaggaaaa gaactattgt ggaagaactg
     3841 ctggctgatt ctgaattcag aagatacaac cgaaggaagt actcagagat catggctgaa
     3901 aaagcagcaa atgcagcagg aaaaaagttc cgaaagaaga agaaatttcg caattaagat
     3961 ttaccaagca aactgcaaca ttttacattg ctcctttatt tacttattaa agacgtttgg
     4021 aaaactaana aaaaaaaaaa aaaaaactcg acggtaccgc gggcccggga tccaccggat
     4081 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     4141 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     4201 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4261 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     4321 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4381 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4441 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4501 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4561 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4621 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4681 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4741 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     4801 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4861 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     4921 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4981 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5041 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5101 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5161 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     5221 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     5281 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5341 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5401 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5461 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     5521 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5581 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5641 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5701 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5761 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5821 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5881 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5941 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6001 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6061 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     6121 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     6181 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     6241 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     6301 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     6361 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     6421 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     6481 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     6541 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6601 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6661 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6721 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6781 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6841 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6901 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6961 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7021 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7081 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7141 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7201 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7261 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7321 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     7381 ctgtggataa ccgtattacc gccatgcat
//
