LOCUS       sequence-413630-        5280 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 5280)
  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 5280)
  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 54kDa
FEATURES             Location/Qualifiers
     source          1..5280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(276..864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(356..375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1193..1240
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1265..1284
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1472..2266)
                     /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             complement(1472..2266)
                     /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(1583..1602)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2193..2212
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2301..2497)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(2309..2519)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2315..2450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2543..2563
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2588..2692)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2719..3174
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2806..2825)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3016..3037
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3192..3211)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     terminator      complement(3237..3284)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3280..3298
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3351..3368)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(join(4068..4778,4779..4781))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(4068..4778,4779..4781))
                     /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(4110..4131)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4459..4478
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4718..4739
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(4874..5077)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4903..4923)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(5078..5280,1..101)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
       61 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
      121 tatgaactaa tgaccccgta attgattact attaataact aatgcatggc ggtaatacgg
      181 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
      241 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
      301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
      721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1021 cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg gccttcaccc
     1081 gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt
     1141 ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac
     1201 gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc
     1261 ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa gaactccagc
     1321 atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc
     1381 aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct
     1441 tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga
     1501 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
     1561 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
     1621 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
     1681 tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg
     1741 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
     1801 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     1861 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     1921 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     1981 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     2041 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg
     2101 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     2161 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     2221 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc
     2281 atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     2341 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
     2401 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
     2461 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     2521 ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg gttctttccg
     2581 cctcaggact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2641 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     2701 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2761 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2821 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2881 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2941 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3001 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3061 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3121 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttacgcgt
     3181 taagatacat tgatgagttt ggacaaacca catccggata tagttcctcc tttcagcaaa
     3241 aaacccctca agacccgttt agaggcccca aggggttatg ctagttattg ctcagcggtg
     3301 gcagcagcca actcagcttc ctttcgggct ttgttagcag ccggatctca gtggtggtgg
     3361 tggtggtgct cgagttattt tggtgcttga gcatcattta gctttttagc ttctgctaaa
     3421 aggttagcgc tttggcttgg gtcatctttt aggctttgga tgaaaccatt gcgttgttct
     3481 tcgtttaagt taggtaaatg taagatttca tagaaagcat tttgttgttc tttgttgaat
     3541 ttgttatccg ctttcggtgc ttgagattca tttaactttt tagcttctga caataggtta
     3601 gcactttggc ttgggtcatc ttttaagctt tggatgaaac cattgcgttg ttcttcgttt
     3661 aagttaggca tattcaagat ttcatagaaa gcattttgtt gttctttgtt gaaattgtta
     3721 tcagctttcg gtgcttgaga ttcgtttaat tttttagctt cacctaaaac gttagtgctt
     3781 tggcttgggt cgtctttaag actttgaatg aagccgttac gttgcgcttc gtttaagtta
     3841 ggcatgttca agatttcata gaaggcgctt tgttgatctt tgttgaagtt attttgttgc
     3901 gcatcagctt ttggagcttg agagtcatta agtttttgag cttcacctaa aacgttagca
     3961 ctttggcttg gatcatcttt aaggctttgg ataaaaccat tgcgttgatc agcatttaag
     4021 ttaggcatat ttaagacttg ataaaaagca ttttgttgag caagcttctt gtacagctcg
     4081 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     4141 ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc
     4201 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg
     4261 ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg
     4321 gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
     4381 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     4441 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     4501 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     4561 tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg
     4621 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc
     4681 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     4741 atgggcacca ccccggtgaa cagctcctcg cccttgctca cgctgccccc gcctccaagt
     4801 tcaagttctt caagtttttt ttgaagcatg gtggcgaccg gtagcgctag cggatctgac
     4861 ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     4921 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     4981 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     5041 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     5101 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     5161 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     5221 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
//
