LOCUS       sequence-400101-        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: M_NLS-mutNLS V5-Twist
FEATURES             Location/Qualifiers
     source          1..4628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        310..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     464..484
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      552..561
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             585..626
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1256..1282
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1295..1312
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1365..1383)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1379..1426
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1452..1471
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1489..1944)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1626..1647)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1838..1857
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1971..2075
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2077..2434
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2077..2268
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2100..2120)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2285..2420
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2347..2366
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2469..3263
                     /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             2469..3263
                     /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(2523..2542)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3133..3152
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3451..3470)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3495..3542
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3871..4459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4360..4379
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccacgc
       61 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      121 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      181 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcactatgcc
      241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      301 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      481 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      541 cgctaccggt cgccaccatg ggcccaaaaa agaaagccaa agttggaaag cctattccta
      601 atcctcttct aggtctagat tctactatgc aggacgtgtc cagctcgcca gtctctccgg
      661 ccgacgacag cctgagcaac agcgaggagg agccggaccg gcagcagccg gcgagcggca
      721 agcgcggggc tcgcaggaga cgcagcagtc ggcgcagcgc gggcggcagc gcggggcccg
      781 gcggggccac gggcgggggc atcggaggcg gcgacgagcc aggcagcccg gcccagggca
      841 ggcgcggcaa gaaatctgcg ggcggaggcg gcggcggcgg cgcgggcgga ggtggtggcg
      901 gcggcggcgg cagcagcagc gggggcggga gcccgcagtc gtacgaggag ctgcagaccc
      961 agcgggtcat ggctaacgtg cgggagcgcc agcgcacgca gtcgctgaac gaggcgttcg
     1021 ccgccctgcg caagatcatc cccacgctgc cctcggacaa gctgagcaag attcagaccc
     1081 tcaaactggc ggccaggtac atcgacttcc tgtaccaggt cctgcagagc gacgagctgg
     1141 actccaagat ggcaagctgc agctatgtgg cccacgagcg gctcagctac gccttctccg
     1201 tctggaggat ggagggggcc tggtccatgt ccgcgtccca ctaggtcgac ccgggtatcc
     1261 gtatgatgtg cctgactacg catgatatct cgagcaccac caccaccacc actgagatcc
     1321 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     1381 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1441 tatatccgga tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     1501 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1561 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1621 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1681 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     1741 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1801 cttgacgggg aaagccggcg aacgtggcga ggaaggaagg gaagaaagcg aaaggagcgg
     1861 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1921 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1981 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2041 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     2101 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2161 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2221 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2281 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2341 tttttttatt tatgcagagg ccgaggccgc ctcggcccct gagctattcc agaagtagtg
     2401 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     2461 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     2521 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     2581 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     2641 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     2701 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     2761 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     2821 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     2881 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     2941 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3001 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3061 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3121 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3181 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3241 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3301 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     3361 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     3421 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     3481 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     3541 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     3601 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     3661 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     3721 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3781 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3841 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3901 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3961 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4021 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4081 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4141 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4201 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4261 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4321 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4381 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4441 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4501 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4561 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     4621 atatggag
//
