LOCUS       sequence-400569-        5736 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 5736)
  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 5736)
  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_NES 67kDa
FEATURES             Location/Qualifiers
     source          1..5736
                     /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"
     CDS             join(616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGNGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2410..2427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2480..2498)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2494..2541
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2567..2586
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2604..3059)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2741..2762)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2953..2972
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3086..3190
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3192..3549
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3192..3383
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3215..3235)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3400..3535
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3584..4378
                     /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             3584..4378
                     /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(3638..3657)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4248..4267
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4566..4585)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4610..4657
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4986..5574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5475..5494
                     /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 caccatgctt tatcctgaga gacttagaag aattcttact
      601 ggaggcgggg gcagcgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcaacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaaga agcttgcaca gcaaaacgct ttctatcaag tactaaacat gccaaacttg
     1381 aatgcagatc aacgtaacgg tttcatccaa agtttgaagg atgatcctag tcaatctgcc
     1441 aacgtgttgg gtgaagctca aaagttgaat gacagtcaag caccaaaggc agatgctcag
     1501 caaaataact ttaacaaaga tcagcaatct gcattctacg agattcttaa tatgcctaat
     1561 ttaaatgagg cgcaacgtaa tggttttatt caaagcctta aggatgatcc atctcaatcc
     1621 acaaacgtcc tgggagaagc aaagaaacta aatgaatcac aagcccctaa ggcagataac
     1681 aactttaata aagaacaaca aaacgcattc tatgaaatcc taaacatgcc taatctgaac
     1741 gaggaacaaa ggaatggctt tatacaatcc ttaaaggatg acccttccca gtctgctaat
     1801 ttattatcag aggctaagaa gttaaacgaa tctcaggctc caaaggccga caataagacg
     1861 cgttttaaca aggagcaaca gaacgctttt tatgaaatct tgcatttacc caacttgaac
     1921 gaagagcaaa gaaatggttt tatacaatct ttaaaggatg atccatcaca aagtgctaac
     1981 cttttagctg aagctaaaaa attaaatgat gcacaagctc caaaggcaga taacaaagtc
     2041 gactttaata aggcacagca aaacgctttc tatcaagtct taaacatgcc aaatctgaat
     2101 gctgatcaaa gaaacggttt catccaatct ttgaaagacg atcctagcca gtccgcaaat
     2161 gtacttggag aagcgcaaaa actgaacgat agccaagctc ctaaagcaga tgcacaacaa
     2221 aacaatttta ataaagatca acaatcagct ttctacgaaa ttctaaacat gccaaacctg
     2281 aatgaagcac aaagaaacgg gtttatccaa tccttaaaag atgacccaag ccaatctacg
     2341 aatgttttgg gagaagctaa gaaacttaat gaatctcaag caccaaaggc cgacaacaat
     2401 taactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
     2461 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     2521 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatgtgg tttgtccaaa
     2581 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     2641 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2701 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2761 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2821 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2881 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2941 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3001 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3061 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3121 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3181 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3241 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3301 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3361 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3421 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3481 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3541 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3601 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3661 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3721 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     3781 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3841 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3901 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3961 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4021 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4081 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4141 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4201 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4261 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4321 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4381 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4441 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4501 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     4561 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4621 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     4681 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4741 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     4801 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     4861 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4921 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4981 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5041 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5101 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5161 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5221 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5281 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5341 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5401 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5461 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5521 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5581 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5641 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     5701 gtaatcaatt acggggtcat tagttcatag cccata
//
