LOCUS       sequence-399851-        4604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Twist 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: Control V5-Twist
FEATURES             Location/Qualifiers
     source          1..4604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      573..582
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             582..623
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1253..1279
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1292..1309
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1362..1380)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1376..1423
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1449..1468
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1486..1941)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1623..1644)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1835..1854
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1968..2072
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2074..2431
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2074..2265
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2097..2117)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2282..2417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2344..2363
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2466..3260
                     /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             2466..3260
                     /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(2520..2539)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3130..3149
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3448..3467)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3492..3539
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3868..4456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4357..4376
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccacg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcactatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      541 gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat gggaaagcct attcctaatc
      601 ctcttctagg tctagattct actatgcagg acgtgtccag ctcgccagtc tctccggccg
      661 acgacagcct gagcaacagc gaggaggagc cggaccggca gcagccggcg agcggcaagc
      721 gcggggctcg caagagacgc agcagtcggc gcagcgcggg cggcagcgcg gggcccggcg
      781 gggccacggg cgggggcatc ggaggcggcg acgagccagg cagcccggcc cagggcaagc
      841 gcggcaagaa atctgcgggc ggaggcggcg gcggcggcgc gggcggaggt ggtggcggcg
      901 gcggcggcag cagcagcggg ggcgggagcc cgcagtcgta cgaggagctg cagacccagc
      961 gggtcatggc taacgtgcgg gagcgccagc gcacgcagtc gctgaacgag gcgttcgccg
     1021 ccctgcgcaa gatcatcccc acgctgccct cggacaagct gagcaagatt cagaccctca
     1081 aactggcggc caggtacatc gacttcctgt accaggtcct gcagagcgac gagctggact
     1141 ccaagatggc aagctgcagc tatgtggccc acgagcggct cagctacgcc ttctccgtct
     1201 ggaggatgga gggggcctgg tccatgtccg cgtcccacta ggtcgacccg ggtatccgta
     1261 tgatgtgcct gactacgcat gatatctcga gcaccaccac caccaccact gagatccggc
     1321 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     1381 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     1441 atccggatgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     1501 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1561 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1621 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1681 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     1741 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     1801 gacggggaaa gccggcgaac gtggcgagga aggaagggaa gaaagcgaaa ggagcgggcg
     1861 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1921 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1981 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2041 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     2101 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2161 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2221 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2281 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2341 ttttatttat gcagaggccg aggccgcctc ggcccctgag ctattccaga agtagtgagg
     2401 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     2461 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2521 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     2581 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     2641 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     2701 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     2761 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     2821 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     2881 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     2941 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     3001 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3061 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3121 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3181 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3241 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     3301 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3361 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3421 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     3481 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     3541 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     3601 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     3661 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     3721 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3781 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3841 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3901 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3961 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4021 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4081 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4141 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4201 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4261 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4321 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4381 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4441 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4501 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4561 cgccatgcat tagttattaa tagtaatcaa ttacggggtc atta
//
