LOCUS       sequence-353791-        8469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses FUS wildtype tagged to mVenus and mCherry in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8469)
  AUTHORS   Wan Q, Mouton SN, Veenhoff LM, Boersma AJ
  TITLE     A FRET-based method for monitoring structural transitions in protein
            self-organization.
  JOURNAL   Cell Rep Methods. 2022 Mar 28;2(3):100184. doi: 
            10.1016/j.crmeth.2022.100184. eCollection 2022 Mar 28.
  PUBMED    35475219
REFERENCE   2  (bases 1 to 8469)
  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"; journalName: "Cell Rep 
            Methods."; date: "2022-03-28"; volume: "2(3):100184. doi"; pages: " 
            10.1016/j.crmeth.2022.100184. eCollection 2022 Mar 28"
COMMENT     Sequence Label: pFUS-WT-VC
FEATURES             Location/Qualifiers
     source          1..8469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        876..1255
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1256..1467
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1418..1438
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1464..1488
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1628..1760
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1743..1762
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1805..1823
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1843..1869
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2250..2261
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             join(3487..3489,3490..4002)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(3529..3550)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3790..3809)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4137..4158
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4298..4316)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4566..4585
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4911..5032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4922..4941)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4976..4995
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      5218..5673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5305..5324)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5515..5536
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5790..6147
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5790..5981
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5813..5833)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5998..6133
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6198..6992
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6252..6271)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6862..6881
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7056..7104
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(7145..7164)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7264..7286
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7324..7342)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7410..7514
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7515..7583,7584..8375)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7733..7752)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatggctcg acagatcttc aatattggcc attagccata ttattcattg gttatatagc
      781 ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat
      841 ttatattggc tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta
      901 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      961 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     1021 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     1081 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     1141 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     1201 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     1261 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
     1321 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     1381 aaaatgtcgt aacaactgcg atcgcccgcc ccgttgacgc aaatgggcgg taggcgtgta
     1441 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcac tagaagcttt
     1501 attgcggtag tttatcacag ttaaattgct aacgcagtca gtgcttctga cacaacagtc
     1561 tcgaacttaa gctgcagtga ctctcttaag gtagccttgc agaagttggt cgtgaggcac
     1621 tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
     1681 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
     1741 ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag
     1801 tacttaatac gactcactat aggctagcct cgagccacca tgtacccata cgacgtccca
     1861 gactacgcta tggcctcaaa cgattatacc caacaagcaa cccaaagcta tggggcctac
     1921 cccacccagc ccgggcaggg ctattcccag cagagcagtc agccctacgg acagcagagt
     1981 tacagtggtt atagccagtc cacggacact tcaggctatg gccagagcag ctattcttct
     2041 tatggccaga gccagaacac aggctatgga actcagtcaa ctccccaggg atatggctcg
     2101 actggcggct atggcagtag ccagagctcc caatcgtctt acgggcagca gtcctcctat
     2161 cctggctatg gccagcagcc agctcccagc agcacctcgg gaagttacgg tagcagttct
     2221 cagagcagca gctatgggca gccccagagt gggagctaca gccagcagcc tagctatggt
     2281 ggacagcagc aaagctatgg acagcagcaa agctataatc cccctcaggg ctatggacag
     2341 cagaaccagt acaacagcag cagtggtggt ggaggtggag gtggaggtgg aggtaactat
     2401 ggccaagatc aatcctccat gagtagtggt ggtggcagtg gtggcggtta tggcaatcaa
     2461 gaccagagtg gtggaggtgg cagcggtggc tatggacagc aggaccgtgg aggccgcggc
     2521 aggggtggca gtggtggcgg cggcggcggc ggcggtggtg gttacaaccg cagcagtggt
     2581 ggctatgaac ccagaggtcg tggaggtggc cgtggaggca gaggtggcat gggcggaagt
     2641 gaccgtggtg gcttcaataa atttggtggc cctcgggacc aaggatcacg tcatgactcc
     2701 gaacaggata attcagacaa caacaccatc tttgtgcaag gcctgggtga gaatgttaca
     2761 attgagtctg tggctgatta cttcaagcag attggtatta ttaagacaaa caagaaaacg
     2821 ggacagccca tgattaattt gtacacagac agggaaactg gcaagctgaa gggagaggca
     2881 acggtctctt ttgatgaccc accttcagct aaagcagcta ttgactggtt tgatggtaaa
     2941 gaattctccg gaaatcctat caaggtctca tttgctactc gccgggcaga ctttaatcgg
     3001 ggtggtggca atggtcgtgg aggccgaggg cgaggaggac ccatgggccg tggaggctat
     3061 ggaggtggtg gcagtggtgg tggtggccga ggaggatttc ccagtggagg tggtggcggt
     3121 ggaggacagc agcgagctgg tgactggaag tgtcctaatc ccacctgtga gaatatgaac
     3181 ttctcttgga ggaatgaatg caaccagtgt aaggccccta aaccagatgg cccaggaggg
     3241 ggaccaggtg gctctcacat ggggggtaac tacggggatg atcgtcgtgg tggcagagga
     3301 ggctatgatc gaggcggcta ccggggccgc ggcggggacc gtggaggctt ccgagggggc
     3361 cggggtggtg gggacagagg tggctttggc cctggcaaga tggattccag gggtgagcac
     3421 agacaggatc gcagggagag gccgtatgga gctcccggca gcgctggatc tgccgcaggc
     3481 tcccttgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3541 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     3601 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
     3661 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
     3721 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3781 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3841 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3901 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     3961 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     4021 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4081 cactacctga gctaccagtc caaactgagc aaagacccca acgagaagcg cgatcacatg
     4141 gtcctgctgg agttcgtgac cgccgcctcc ggagataaca tggccatcat caaggagttc
     4201 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     4261 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     4321 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     4381 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     4441 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     4501 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     4561 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     4621 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     4681 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     4741 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     4801 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatgtaatc tagagtcgac
     4861 ccgggcggcc gcttcccttt agtgagggtt aatgcttcga gcagacatga taagatacat
     4921 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4981 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     5041 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa
     5101 gtaaaacctc tacaaatgtg gtaaaatccg ataaggatcg atccgggctg gcgtaatagc
     5161 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     5221 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5281 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5341 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5401 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5461 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5521 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5581 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5641 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     5701 tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc
     5761 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     5821 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5881 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5941 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     6001 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6061 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     6121 ttttttggag gcctaggctt ttgcaaaaag cttgattctt ctgacacaac agtctcgaac
     6181 ttaaggctag agccaccatg attgaacaag atggattgca cgcaggttct ccggccgctt
     6241 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     6301 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     6361 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     6421 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     6481 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     6541 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     6601 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     6661 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     6721 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     6781 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     6841 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     6901 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     6961 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     7021 ccaagcgacg cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac
     7081 atctgtgtgt tggttttttg tgtgaatcga tagcgataag gatccgcgta tggtgcactc
     7141 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     7201 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     7261 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     7321 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     7381 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     7441 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     7501 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     7561 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     7621 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     7681 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     7741 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     7801 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     7861 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     7921 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     7981 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     8041 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     8101 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     8161 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     8221 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     8281 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     8341 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     8401 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     8461 ttgataatc
//
