LOCUS       sequence-400102-        4628 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 4628)
  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 4628)
  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-mutNLS V5-Twist
FEATURES             Location/Qualifiers
     source          1..4628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..313
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        314..517
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     468..488
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      556..565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             589..630
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1260..1286
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1299..1316
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1369..1387)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1383..1430
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1456..1475
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1493..1948)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1630..1651)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1842..1861
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1975..2079
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2081..2438
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2081..2272
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2104..2124)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2289..2424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2351..2370
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2473..3267
                     /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             2473..3267
                     /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(2527..2546)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3137..3156
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3455..3474)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3499..3546
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3875..4463
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4364..4383
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
       61 acgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      121 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      181 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcacta
      241 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      301 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      361 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      421 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      481 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
      541 ctagcgctac cggtcgccac catgggccca aaaaaggcca gaaaagttgg aaagcctatt
      601 cctaatcctc ttctaggtct agattctact atgcaggacg tgtccagctc gccagtctct
      661 ccggccgacg acagcctgag caacagcgag gaggagccgg accggcagca gccggcgagc
      721 ggcaagcgcg gggctcgcag gagacgcagc agtcggcgca gcgcgggcgg cagcgcgggg
      781 cccggcgggg ccacgggcgg gggcatcgga ggcggcgacg agccaggcag cccggcccag
      841 ggcaggcgcg gcaagaaatc tgcgggcgga ggcggcggcg gcggcgcggg cggaggtggt
      901 ggcggcggcg gcggcagcag cagcgggggc gggagcccgc agtcgtacga ggagctgcag
      961 acccagcggg tcatggctaa cgtgcgggag cgccagcgca cgcagtcgct gaacgaggcg
     1021 ttcgccgccc tgcgcaagat catccccacg ctgccctcgg acaagctgag caagattcag
     1081 accctcaaac tggcggccag gtacatcgac ttcctgtacc aggtcctgca gagcgacgag
     1141 ctggactcca agatggcaag ctgcagctat gtggcccacg agcggctcag ctacgccttc
     1201 tccgtctgga ggatggaggg ggcctggtcc atgtccgcgt cccactaggt cgacccgggt
     1261 atccgtatga tgtgcctgac tacgcatgat atctcgagca ccaccaccac caccactgag
     1321 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     1381 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     1441 gaactatatc cggatgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     1501 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     1561 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     1621 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     1681 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     1741 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     1801 agagcttgac ggggaaagcc ggcgaacgtg gcgaggaagg aagggaagaa agcgaaagga
     1861 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     1921 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     1981 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2041 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     2101 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2161 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2221 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2281 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2341 taattttttt tatttatgca gaggccgagg ccgcctcggc ccctgagcta ttccagaagt
     2401 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     2461 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     2521 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     2581 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     2641 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     2701 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     2761 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     2821 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     2881 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     2941 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3001 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3061 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3121 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3181 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3241 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     3301 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     3361 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     3421 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     3481 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     3541 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     3601 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     3661 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     3721 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3781 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3841 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3901 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3961 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4021 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4081 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4141 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4201 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4261 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4321 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4381 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4441 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4501 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4561 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     4621 ccatatat
//
