LOCUS       sequence-399869-        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: mut GBP2 V5-Twist
FEATURES             Location/Qualifiers
     source          1..4604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        31..334
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        335..538
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     489..509
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             586..627
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1257..1283
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1296..1313
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1366..1384)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1380..1427
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1453..1472
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1490..1945)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1627..1648)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1839..1858
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1972..2076
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2078..2435
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2078..2269
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2101..2121)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2286..2421
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2348..2367
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2470..3264
                     /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             2470..3264
                     /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(2524..2543)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3134..3153
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3452..3471)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3496..3543
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3872..4460
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4361..4380
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
       61 tggctgaccg cccaacgacc cacgcccatt gacgtcaata atgacgtatg ttcccatagt
      121 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      181 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      241 taaatggccc gcctggcact atgcccagta catgacctta tgggactttc ctacttggca
      301 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      361 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      421 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      481 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      541 tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatgggaaa gcctattcct
      601 aatcctcttc taggtctaga ttctactatg caggacgtgt ccagctcgcc agtctctccg
      661 gccgacgaca gcctgagcaa cagcgaggag gagccggacc ggcagcagcc ggcgagcggc
      721 aagcgcgggg ctcgcaagag acgcagcagt cggcgcagcg cgggcggcag cgcggggccc
      781 ggcggggcca cgggcggggg catcggaggc ggcgacgagc caggcagccc ggcccagggc
      841 aagcgcggca agaaatctgc gggcggaggc ggcggcggcg gcgcgggcgg aggtggtggc
      901 ggcggcggcg gcagcagcag cgggggcggg agcccgcagt cggaggagga gctgcagacc
      961 cagcgggtca tggctaacgt gcgggagcgc cagcgcacgc agtcgctgaa cgaggcgttc
     1021 gccgccctgc gcaagatcat ccccacgctg ccctcggaca agctgagcaa gattcagacc
     1081 ctcaaactgg cggccaggta catcgacttc ctgtaccagg tcctgcagag cgacgagctg
     1141 gactccaaga tggcaagctg cagctatgtg gcccacgagc ggctcagcta cgccttctcc
     1201 gtctggagga tggagggggc ctggtccatg tccgcgtccc actaggtcga cccgggtatc
     1261 cgtatgatgt gcctgactac gcatgatatc tcgagcacca ccaccaccac cactgagatc
     1321 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     1381 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     1441 ctatatccgg atgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     1501 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     1561 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     1621 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     1681 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     1741 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     1801 gcttgacggg gaaagccggc gaacgtggcg aggaaggaag ggaagaaagc gaaaggagcg
     1861 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     1921 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     1981 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2041 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     2101 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2161 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2221 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2281 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2341 ttttttttat ttatgcagag gccgaggccg cctcggcccc tgagctattc cagaagtagt
     2401 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     2461 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     2521 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     2581 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     2641 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     2701 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     2761 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     2821 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     2881 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     2941 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     3001 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3061 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3121 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3181 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3241 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3301 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3361 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3421 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     3481 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     3541 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     3601 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     3661 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     3721 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3781 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3841 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3901 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3961 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4021 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4081 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4141 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4201 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4261 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4321 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4381 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4441 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4501 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4561 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtc
//
