LOCUS       sequence-399088-        5235 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 5235)
  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 5235)
  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: Diffusive 54kDa
FEATURES             Location/Qualifiers
     source          1..5235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        13..316
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        317..520
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     471..491
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      559..568
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(565..567,568..570,571..1281)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(565..567,568..570,571..1281)
                     /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(610..631)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(871..890)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1218..1239
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1981..1998
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2051..2069)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2065..2112
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2138..2157
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2175..2630)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2312..2333)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2524..2543
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2657..2761
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2786..2806)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2830..3040
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2852..3048
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2899..3034
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3083..3877
                     /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             3083..3877
                     /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(3137..3156)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3747..3766
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4065..4084)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4109..4156
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4485..5073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4974..4993
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
       61 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      121 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      181 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      241 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      301 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      361 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      421 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      481 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      541 ccgctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
      601 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      661 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      721 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
      781 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      841 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      901 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      961 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1021 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     1081 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1141 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1201 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1261 ggcatggacg agctgtacaa gaagcttgct caacaaaatg ctttttatca agtcttaaat
     1321 atgcctaact taaatgctga tcaacgcaat ggttttatcc aaagccttaa agatgatcca
     1381 agccaaagtg ctaacgtttt aggtgaagct caaaaactta atgactctca agctccaaaa
     1441 gctgatgcgc aacaaaataa cttcaacaaa gatcaacaaa gcgccttcta tgaaatcttg
     1501 aacatgccta acttaaacga agcgcaacgt aacggcttca ttcaaagtct taaagacgac
     1561 ccaagccaaa gcactaacgt tttaggtgaa gctaaaaaat taaacgaatc tcaagcaccg
     1621 aaagctgata acaatttcaa caaagaacaa caaaatgctt tctatgaaat cttgaatatg
     1681 cctaacttaa acgaagaaca acgcaatggt ttcatccaaa gcttaaaaga tgacccaagc
     1741 caaagtgcta acctattgtc agaagctaaa aagttaaatg aatctcaagc accgaaagcg
     1801 gataacaaat tcaacaaaga acaacaaaat gctttctatg aaatcttaca tttacctaac
     1861 ttaaacgaag aacaacgcaa tggtttcatc caaagcctaa aagatgaccc aagccaaagc
     1921 gctaaccttt tagcagaagc taaaaagcta aatgatgctc aagcaccaaa ataactcgag
     1981 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     2041 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     2101 aggggttttt tgctgaaagg aggaactata tccggatgtg gtttgtccaa actcatcaat
     2161 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2221 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2281 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2341 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2401 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2461 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2521 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     2581 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     2641 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2701 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2761 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2821 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2881 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2941 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3001 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3061 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3121 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3181 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3241 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3301 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3361 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3421 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3481 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3541 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3601 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3661 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3721 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3781 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3841 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3901 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3961 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4021 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4081 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4141 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4201 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4261 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4321 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4381 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4441 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4501 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4561 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4621 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4681 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4741 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4801 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4861 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4921 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4981 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5041 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5101 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5161 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     5221 agttcatagc ccata
//
