LOCUS       sequence-133053-        5585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively nuclear variant of a very strong, photocaged NES 
            (AsLOV2-NES 27) for blocking endogenous, CRM-1-dependent nuclear 
            export.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5585)
  AUTHORS   Niopek D, Wehler P, Roensch J, Eils R, Di Ventura B
  TITLE     Optogenetic control of nuclear protein export.
  JOURNAL   Nat Commun. 2016 Feb 8;7:10624. doi: 10.1038/ncomms10624.
  PUBMED    26853913
REFERENCE   2  (bases 1 to 5585)
  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: "Nat 
            Commun."; date: "2016-02-8"; volume: "7:10624. doi"; pages: " 
            10.1038/ncomms10624"
COMMENT     Sequence Label: H2B-GFP-AsLOV2-NES27 (pDN158)
FEATURES             Location/Qualifiers
     source          1..5585
                     /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"
     CDS             657..1034
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1047..1056
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1053..1055,1056..1058,1059..1769)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1053..1055,1056..1058,1059..1769)
                     /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(1098..1119)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1359..1378)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1706..1727
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2373..2494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2410..2429)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2464..2483
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2501..2956)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2638..2659)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2850..2869
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2983..3087
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3089..3446
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3089..3280
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3112..3132)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3297..3432
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3481..4275
                     /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             3481..4275
                     /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(3535..3554)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4145..4164
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4463..4482)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4507..4554
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4883..5471
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5372..5391
                     /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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gccaccatgc
      661 cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg gtgactaagg
      721 cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat tccatctatg
      781 tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag gccatgggca
      841 tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct tcccgcctgg
      901 cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc gtgcgcctgc
      961 tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc atcaccaagt
     1021 acaccagcgc taaggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca
     1081 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1141 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1201 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1261 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1321 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1381 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1441 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1501 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1561 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1621 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1681 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1741 tcactctcgg catggacgag ctgtacaagg gtggatctgg aggttcaggt ggaagtttgg
     1801 ctactacact tgaacgtatt gagaagaact ttgtcattac tgacccaaga ttgccagata
     1861 atcccattat attcgcgtcc gatagtttct tgcagttgac agaatatagc cgtgaagaaa
     1921 ttttgggaag aaactgcagg tttctacaag gtcctgaaac tgatcgcgcg acagtgagaa
     1981 aaattagaga tgccatagat aaccaaacag aggtcactgt tcagctgatt aattatacaa
     2041 agagtggtaa aaagttctgg aacctctttc acttgcagcc tatgcgagat cagaagggag
     2101 atgtccagta ctttattggg gttcagttgg atggaactga gcatgtccga gatgctgccg
     2161 agagagaggg agtcatgctg attaagaaaa ctgcagaaaa tattgaggag ctggccaagg
     2221 agctggccga cctgaacctg gactagctcg agcggccgcg actctagatc ataatcagcc
     2281 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2341 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2401 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2461 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     2521 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2581 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2641 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2701 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     2761 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     2821 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     2881 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     2941 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3001 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3061 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3121 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3181 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3241 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3301 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3361 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3421 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     3481 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3541 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3601 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3661 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3721 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3781 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3841 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3901 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3961 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4021 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4081 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4141 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4201 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4261 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4321 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4381 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4441 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4501 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     4561 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4621 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4681 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     4741 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4801 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4861 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4921 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4981 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5041 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5101 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5161 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5221 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5281 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5341 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5401 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5461 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5521 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     5581 tgcat
//
