LOCUS       sequence-400068-        5178 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 5178)
  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 5178)
  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_NES 47kDa
FEATURES             Location/Qualifiers
     source          1..5178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        21..324
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        325..528
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     479..499
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(621..623,624..1334)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(621..623,624..1334)
                     /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(663..684)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(924..943)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1271..1292
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1860..1877
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1930..1948)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1944..1991
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2017..2036
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2054..2509)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2191..2212)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2403..2422
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2536..2640
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2642..2999
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2642..2833
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2665..2685)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2850..2985
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3034..3828
                     /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             3034..3828
                     /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(3088..3107)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3698..3717
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4016..4035)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4060..4107
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4436..5024
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4925..4944
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
       61 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      121 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      181 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      241 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      301 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      361 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      421 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      481 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      541 cgtcagatcc gctagcgcta ccggtcgcca ccatgcttca aaaaaaactt gaagaacttg
      601 aacttggagg cgggggcagc gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      661 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      721 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      781 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
      841 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      901 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
      961 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1021 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     1081 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     1141 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1201 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     1261 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1321 acgagctgta caagaagctt gctcaacaaa atgcttttta tcaagtctta aatatgccta
     1381 acttaaatgc tgatcaacgc aatggtttta tccaaagcct taaagatgat ccaagccaaa
     1441 gtgctaacgt tttaggtgaa gctcaaaaac ttaatgactc tcaagctcca aaagctgatg
     1501 cgcaacaaaa taacttcaac aaagatcaac aaagcgcctt ctatgaaatc ttgaacatgc
     1561 ctaacttaaa cgaagcgcaa cgtaacggct tcattcaaag tcttaaagac gacccaagcc
     1621 aaagcactaa cgttttaggt gaagctaaaa aattaaacga atctcaagca ccgaaagctg
     1681 ataacaattt caacaaagaa caacaaaatg ctttctatga aatcttgaat atgcctaact
     1741 taaacgaaga acaacgcaat ggtttcatcc aaagcttaaa agatgaccca agccaaagtg
     1801 ctaacctatt gtcagaagct aaaaagttaa atgaatctca agcaccgaaa taactcgagc
     1861 accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     1921 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     1981 ggggtttttt gctgaaagga ggaactatat ccggatgtgg tttgtccaaa ctcatcaatg
     2041 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2101 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2161 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2221 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2281 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2341 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2401 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2461 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     2521 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2581 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2641 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2701 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2761 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2821 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2881 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2941 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     3001 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3061 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3121 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3181 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     3241 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3301 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3361 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3421 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3481 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3541 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     3601 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3661 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3721 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3781 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     3841 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     3901 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3961 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     4021 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4081 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4141 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     4201 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     4261 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     4321 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4381 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4441 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4501 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4561 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4621 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4681 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4741 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4801 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4861 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4921 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4981 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5041 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5101 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
     5161 acggggtcat tagttcat
//
