LOCUS       sequence-400098-        5726 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 5726)
  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 5726)
  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 67kDa
FEATURES             Location/Qualifiers
     source          1..5726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        24..327
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        328..531
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     482..502
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      570..579
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(618..620,621..1331)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(618..620,621..1331)
                     /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(660..681)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(921..940)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1268..1289
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2412..2429
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2482..2500)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2496..2543
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2569..2588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2606..3061)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2743..2764)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2955..2974
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3088..3192
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3194..3551
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3194..3385
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3217..3237)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3402..3537
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3586..4380
                     /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             3586..4380
                     /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(3640..3659)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4250..4269
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4568..4587)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4612..4659
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4988..5575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5476..5495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
       61 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      121 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      181 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      241 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      301 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      361 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      421 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      481 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
      541 aaccgtcaga tccgctagcg ctaccggtcg ccaccatggg cccaaaaaag gccagaaaag
      601 ttggaggcgg gggcagcgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      661 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      721 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
      781 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
      841 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      901 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
      961 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1021 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1081 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1141 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1201 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     1261 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1321 agctgtacaa gaagcttgca cagcaaaacg ctttctatca agtactaaac atgccaaact
     1381 tgaatgcaga tcaacgtaac ggtttcatcc aaagtttgaa ggatgatcct agtcaatctg
     1441 ccaacgtgtt gggtgaagct caaaagttga atgacagtca agcaccaaag gcagatgctc
     1501 agcaaaataa ctttaacaaa gatcagcaat ctgcattcta cgagattctt aatatgccta
     1561 atttaaatga ggcgcaacgt aatggtttta ttcaaagcct taaggatgat ccatctcaat
     1621 ccacaaacgt cctgggagaa gcaaagaaac taaatgaatc acaagcccct aaggcagata
     1681 acaactttaa taaagaacaa caaaacgcat tctatgaaat cctaaacatg cctaatctga
     1741 acgaggaaca aaggaatggc tttatacaat ccttaaagga tgacccttcc cagtctgcta
     1801 atttattatc agaggctaag aagttaaacg aatctcaggc tccaaaggcc gacaataaga
     1861 cgcgttttaa caaggagcaa cagaacgctt tttatgaaat cttgcattta cccaacttga
     1921 acgaagagca aagaaatggt tttatacaat ctttaaagga tgatccatca caaagtgcta
     1981 accttttagc tgaagctaaa aaattaaatg atgcacaagc tccaaaggca gataacaaag
     2041 tcgactttaa taaggcacag caaaacgctt tctatcaagt cttaaacatg ccaaatctga
     2101 atgctgatca aagaaacggt ttcatccaat ctttgaaaga cgatcctagc cagtccgcaa
     2161 atgtacttgg agaagcgcaa aaactgaacg atagccaagc tcctaaagca gatgcacaac
     2221 aaaacaattt taataaagat caacaatcag ctttctacga aattctaaac atgccaaacc
     2281 tgaatgaagc acaaagaaac gggtttatcc aatccttaaa agatgaccca agccaatcta
     2341 cgaatgtttt gggagaagct aagaaactta atgaatctca agcaccaaag gccgacaaca
     2401 attaactcga gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg
     2461 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     2521 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatgt ggtttgtcca
     2581 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     2641 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2701 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2761 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2821 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2881 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2941 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3001 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3061 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3121 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3181 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3241 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3301 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3361 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3421 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3481 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3541 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3601 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3661 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3721 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3781 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3841 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3901 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3961 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4021 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4081 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4141 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4201 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4261 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4321 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4381 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4441 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4501 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     4561 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4621 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4681 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4741 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4801 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4861 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4921 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4981 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaccacc
     5041 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5101 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5161 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5221 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5281 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5341 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5401 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5461 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5521 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5581 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5641 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     5701 agtaatcaat tacggggtca ttagtt
//
