LOCUS       sequence-400093-        4604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Twist mutant for nucleocytoplasmic transport studies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4604)
  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 4604)
  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: mutNLS V5-Twist
FEATURES             Location/Qualifiers
     source          1..4604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        19..322
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        323..526
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     477..497
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      565..574
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             574..615
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1245..1271
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1284..1301
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1354..1372)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1368..1415
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1441..1460
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1478..1933)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1615..1636)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1827..1846
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1960..2064
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2066..2423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2066..2257
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2089..2109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2274..2409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2336..2355
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2458..3252
                     /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             2458..3252
                     /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(2512..2531)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3122..3141
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3440..3459)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3484..3531
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3860..4448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4349..4368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
       61 caacgaccca cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      121 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      181 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      241 ctggcactat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      301 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      361 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      421 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      481 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      541 tcagatccgc tagcgctacc ggtcgccacc atgggaaagc ctattcctaa tcctcttcta
      601 ggtctagatt ctactatgca ggacgtgtcc agctcgccag tctctccggc cgacgacagc
      661 ctgagcaaca gcgaggagga gccggaccgg cagcagccgg cgagcggcaa gcgcggggct
      721 cgcaggagac gcagcagtcg gcgcagcgcg ggcggcagcg cggggcccgg cggggccacg
      781 ggcgggggca tcggaggcgg cgacgagcca ggcagcccgg cccagggcag gcgcggcaag
      841 aaatctgcgg gcggaggcgg cggcggcggc gcgggcggag gtggtggcgg cggcggcggc
      901 agcagcagcg ggggcgggag cccgcagtcg tacgaggagc tgcagaccca gcgggtcatg
      961 gctaacgtgc gggagcgcca gcgcacgcag tcgctgaacg aggcgttcgc cgccctgcgc
     1021 aagatcatcc ccacgctgcc ctcggacaag ctgagcaaga ttcagaccct caaactggcg
     1081 gccaggtaca tcgacttcct gtaccaggtc ctgcagagcg acgagctgga ctccaagatg
     1141 gcaagctgca gctatgtggc ccacgagcgg ctcagctacg ccttctccgt ctggaggatg
     1201 gagggggcct ggtccatgtc cgcgtcccac taggtcgacc cgggtatccg tatgatgtgc
     1261 ctgactacgc atgatatctc gagcaccacc accaccacca ctgagatccg gctgctaaca
     1321 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     1381 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     1441 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     1501 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     1561 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     1621 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     1681 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     1741 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     1801 aagccggcga acgtggcgag gaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     1861 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     1921 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1981 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2041 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     2101 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2161 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2221 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2281 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2341 atgcagaggc cgaggccgcc tcggcccctg agctattcca gaagtagtga ggaggctttt
     2401 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     2461 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     2521 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     2581 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     2641 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     2701 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     2761 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     2821 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     2881 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     2941 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     3001 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3061 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3121 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3181 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     3241 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     3301 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     3361 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     3421 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     3481 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     3541 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     3601 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     3661 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     3721 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3781 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3841 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3901 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3961 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4021 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4081 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4141 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4201 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4261 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4321 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4381 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4441 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4501 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     4561 attagttatt aatagtaatc aattacgggg tcattagttc atag
//
