LOCUS       sequence-400571-        5733 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 5733)
  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 5733)
  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 67kDa
FEATURES             Location/Qualifiers
     source          1..5733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        20..323
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        324..527
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     478..498
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(620..622,623..1333)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(620..622,623..1333)
                     /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(662..683)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(923..942)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1270..1291
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2414..2431
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2484..2502)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2498..2545
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2571..2590
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2608..3063)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2745..2766)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2957..2976
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3090..3194
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3196..3553
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3196..3387
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3219..3239)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3404..3539
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3588..4382
                     /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             3588..4382
                     /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(3642..3661)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4252..4271
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4570..4589)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4614..4661
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4990..5578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5479..5498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
       61 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      121 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      181 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      241 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      301 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      361 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      421 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      481 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      541 gtcagatccg ctagcgctac cggtcgccac catgcttcaa aaaaaacttg aagaacttga
      601 acttggaggc gggggcagcg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      661 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      721 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      781 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      841 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      901 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      961 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1021 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1081 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1141 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1201 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     1261 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1321 cgagctgtac aagaagcttg cacagcaaaa cgctttctat caagtactaa acatgccaaa
     1381 cttgaatgca gatcaacgta acggtttcat ccaaagtttg aaggatgatc ctagtcaatc
     1441 tgccaacgtg ttgggtgaag ctcaaaagtt gaatgacagt caagcaccaa aggcagatgc
     1501 tcagcaaaat aactttaaca aagatcagca atctgcattc tacgagattc ttaatatgcc
     1561 taatttaaat gaggcgcaac gtaatggttt tattcaaagc cttaaggatg atccatctca
     1621 atccacaaac gtcctgggag aagcaaagaa actaaatgaa tcacaagccc ctaaggcaga
     1681 taacaacttt aataaagaac aacaaaacgc attctatgaa atcctaaaca tgcctaatct
     1741 gaacgaggaa caaaggaatg gctttataca atccttaaag gatgaccctt cccagtctgc
     1801 taatttatta tcagaggcta agaagttaaa cgaatctcag gctccaaagg ccgacaataa
     1861 gacgcgtttt aacaaggagc aacagaacgc tttttatgaa atcttgcatt tacccaactt
     1921 gaacgaagag caaagaaatg gttttataca atctttaaag gatgatccat cacaaagtgc
     1981 taacctttta gctgaagcta aaaaattaaa tgatgcacaa gctccaaagg cagataacaa
     2041 agtcgacttt aataaggcac agcaaaacgc tttctatcaa gtcttaaaca tgccaaatct
     2101 gaatgctgat caaagaaacg gtttcatcca atctttgaaa gacgatccta gccagtccgc
     2161 aaatgtactt ggagaagcgc aaaaactgaa cgatagccaa gctcctaaag cagatgcaca
     2221 acaaaacaat tttaataaag atcaacaatc agctttctac gaaattctaa acatgccaaa
     2281 cctgaatgaa gcacaaagaa acgggtttat ccaatcctta aaagatgacc caagccaatc
     2341 tacgaatgtt ttgggagaag ctaagaaact taatgaatct caagcaccaa aggccgacaa
     2401 caattaactc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
     2461 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     2521 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat gtggtttgtc
     2581 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     2641 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2701 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2761 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2821 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2881 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2941 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3001 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3061 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3181 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3241 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3301 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3361 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3421 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3481 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3541 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3601 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3661 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3721 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     3781 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3841 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3901 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3961 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4021 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4081 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4141 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4201 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4261 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4321 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4381 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4441 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4501 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     4561 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4621 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4681 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4741 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4801 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     4861 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4921 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4981 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5041 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5101 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5161 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5221 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5281 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5341 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5401 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5461 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5521 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5581 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5641 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     5701 aatagtaatc aattacgggg tcattagttc ata
//
