LOCUS       sequence-400592-        5274 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 5274)
  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 5274)
  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: L_NLS 54kDa
FEATURES             Location/Qualifiers
     source          1..5274
                     /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"
     regulatory      567..576
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(615..617,618..1328)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(615..617,618..1328)
                     /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(657..678)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(918..937)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1265..1286
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2028..2045
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2098..2116)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2112..2159
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2185..2204
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2222..2677)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2359..2380)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2571..2590
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2704..2808
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2833..2853)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2877..3087
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2899..3095
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2946..3081
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3130..3924
                     /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             3130..3924
                     /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(3184..3203)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3794..3813
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4112..4131)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4156..4203
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4532..5120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5021..5040
                     /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 ccatgggccc aaaaaaggcc agaaaagttg
      601 gaggcggggg cagcgtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      661 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      721 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      781 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
      841 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      901 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      961 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1021 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1081 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1141 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1201 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1261 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1321 tgtacaagaa gcttgctcaa caaaatgctt tttatcaagt cttaaatatg cctaacttaa
     1381 atgctgatca acgcaatggt tttatccaaa gccttaaaga tgatccaagc caaagtgcta
     1441 acgttttagg tgaagctcaa aaacttaatg actctcaagc tccaaaagct gatgcgcaac
     1501 aaaataactt caacaaagat caacaaagcg ccttctatga aatcttgaac atgcctaact
     1561 taaacgaagc gcaacgtaac ggcttcattc aaagtcttaa agacgaccca agccaaagca
     1621 ctaacgtttt aggtgaagct aaaaaattaa acgaatctca agcaccgaaa gctgataaca
     1681 atttcaacaa agaacaacaa aatgctttct atgaaatctt gaatatgcct aacttaaacg
     1741 aagaacaacg caatggtttc atccaaagct taaaagatga cccaagccaa agtgctaacc
     1801 tattgtcaga agctaaaaag ttaaatgaat ctcaagcacc gaaagcggat aacaaattca
     1861 acaaagaaca acaaaatgct ttctatgaaa tcttacattt acctaactta aacgaagaac
     1921 aacgcaatgg tttcatccaa agcctaaaag atgacccaag ccaaagcgct aaccttttag
     1981 cagaagctaa aaagctaaat gatgctcaag caccaaaata actcgagcac caccaccacc
     2041 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     2101 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     2161 tgaaaggagg aactatatcc ggatgtggtt tgtccaaact catcaatgta tcttaacgcg
     2221 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2281 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2341 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2401 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2461 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2521 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2581 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2641 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2701 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2761 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     2821 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2881 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2941 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3001 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3061 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     3121 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3181 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3241 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3301 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3361 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3421 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3481 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3541 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3601 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     3661 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3721 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3781 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3841 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3901 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3961 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4021 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4081 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4141 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     4201 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4261 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4321 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     4381 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4441 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4501 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4561 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4621 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4681 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4741 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4801 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4861 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4921 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4981 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5041 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5101 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5161 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5221 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcat
//
