LOCUS       sequence-400588-        5727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nucleocytoplasmic transport probe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5727)
  AUTHORS   Andreu I, Granero-Moya I, Chahare NR, Clein K, Molina-Jordan M, 
            Beedle AEM, Elosegui-Artola A, Abenza JF, Rossetti L, Trepat X, 
            Raveh B, Roca-Cusachs P
  TITLE     Mechanical force application to the nucleus regulates 
            nucleocytoplasmic transport.
  JOURNAL   Nat Cell Biol. 2022 Jun;24(6):896-905. doi: 
            10.1038/s41556-022-00927-7. Epub 2022 Jun 9.
  PUBMED    35681009
REFERENCE   2  (bases 1 to 5727)
  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: 
            "10.1038/s41556-022-00927-7"; journalName: "Nat Cell Biol"; date: 
            "2022-06"; volume: "24"; issue: "6"; pages: "896-905"
COMMENT     Sequence Label: M_NLS 67kDa
FEATURES             Location/Qualifiers
     source          1..5727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        20..323
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        324..527
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     478..498
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      566..575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(614..616,617..1327)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(614..616,617..1327)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(656..677)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(917..936)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1264..1285
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2408..2425
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2478..2496)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2492..2539
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2565..2584
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2602..3057)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2739..2760)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2951..2970
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3084..3188
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3190..3547
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3190..3381
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3213..3233)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3398..3533
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3582..4376
                     /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             3582..4376
                     /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(3636..3655)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4246..4265
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4564..4583)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4608..4655
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4984..5572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5473..5492
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
       61 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      121 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      181 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      241 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      301 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      361 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      421 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      481 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      541 gtcagatccg ctagcgctac cggtcgccac catgggccca aaaaagaaag ccaaagttgg
      601 aggcgggggc agcgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      661 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      721 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      781 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      841 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      901 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      961 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1021 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1081 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1141 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1201 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1261 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1321 gtacaagaag cttgcacagc aaaacgcttt ctatcaagta ctaaacatgc caaacttgaa
     1381 tgcagatcaa cgtaacggtt tcatccaaag tttgaaggat gatcctagtc aatctgccaa
     1441 cgtgttgggt gaagctcaaa agttgaatga cagtcaagca ccaaaggcag atgctcagca
     1501 aaataacttt aacaaagatc agcaatctgc attctacgag attcttaata tgcctaattt
     1561 aaatgaggcg caacgtaatg gttttattca aagccttaag gatgatccat ctcaatccac
     1621 aaacgtcctg ggagaagcaa agaaactaaa tgaatcacaa gcccctaagg cagataacaa
     1681 ctttaataaa gaacaacaaa acgcattcta tgaaatccta aacatgccta atctgaacga
     1741 ggaacaaagg aatggcttta tacaatcctt aaaggatgac ccttcccagt ctgctaattt
     1801 attatcagag gctaagaagt taaacgaatc tcaggctcca aaggccgaca ataagacgcg
     1861 ttttaacaag gagcaacaga acgcttttta tgaaatcttg catttaccca acttgaacga
     1921 agagcaaaga aatggtttta tacaatcttt aaaggatgat ccatcacaaa gtgctaacct
     1981 tttagctgaa gctaaaaaat taaatgatgc acaagctcca aaggcagata acaaagtcga
     2041 ctttaataag gcacagcaaa acgctttcta tcaagtctta aacatgccaa atctgaatgc
     2101 tgatcaaaga aacggtttca tccaatcttt gaaagacgat cctagccagt ccgcaaatgt
     2161 acttggagaa gcgcaaaaac tgaacgatag ccaagctcct aaagcagatg cacaacaaaa
     2221 caattttaat aaagatcaac aatcagcttt ctacgaaatt ctaaacatgc caaacctgaa
     2281 tgaagcacaa agaaacgggt ttatccaatc cttaaaagat gacccaagcc aatctacgaa
     2341 tgttttggga gaagctaaga aacttaatga atctcaagca ccaaaggccg acaacaatta
     2401 actcgagcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
     2461 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     2521 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatgtggtt tgtccaaact
     2581 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2641 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2701 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2761 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2821 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2881 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2941 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3001 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3061 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3121 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3181 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3301 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3361 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3421 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3481 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3541 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3601 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3661 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3721 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     3781 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3841 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3901 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3961 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4021 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4081 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4141 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4201 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4261 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4321 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4381 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4441 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4501 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4561 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4621 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4681 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4741 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4801 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     4861 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4921 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4981 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5041 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5101 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5161 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5221 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5281 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5341 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5401 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5461 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5521 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5581 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5641 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     5701 aatcaattac ggggtcatta gttcata
//
