LOCUS       sequence-372335-        8442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses mVenus and mCherry tagged to FUS wildtype in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8442)
  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 8442)
  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: pVC-FUS-WT
FEATURES             Location/Qualifiers
     source          1..8442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        75..378
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        379..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          751..883
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     866..885
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        928..946
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             966..992
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(993..995,996..998,999..1511)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(1038..1059)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1299..1318)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1646..1667
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1807..1825)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2075..2094
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2744..2755
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    4007..4128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4018..4037)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4072..4091
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4314..4769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4401..4420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4611..4632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4886..5243
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4886..5077
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4909..4929)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5094..5229
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5294..6088
                     /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(5348..5367)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5958..5977
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6152..6200
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6241..6260)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6360..6382
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6420..6438)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6506..6610
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6611..6679,6680..7471)
                     /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(6829..6848)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7642..8230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8131..8150
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
       61 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      121 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      181 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      241 gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      301 cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta
      361 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg
      421 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      481 caccaaaatc aacgggactt tccaaaatgt cgtaacaact gcgatcgccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cactagaagc tttattgcgg tagtttatca cagttaaatt gctaacgcag
      661 tcagtgcttc tgacacaaca gtctcgaact taagctgcag tgactctctt aaggtagcct
      721 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
      781 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
      841 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc
      901 aattacagct cttaaggcta gagtacttaa tacgactcac tataggctag cctcgagcca
      961 ccatgtaccc atacgacgtc ccagactacg ctatggtgag caagggcgag gagctgttca
     1021 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1081 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
     1141 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcctgc
     1201 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1261 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1321 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1381 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1441 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     1501 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1561 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aaactgagca
     1621 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgcctccg
     1681 gagataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     1741 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     1801 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     1861 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     1921 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     1981 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     2041 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     2101 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     2161 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     2221 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     2281 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     2341 ccaccggcgg catgggctcc cttatggcct caaacgatta tacccaacaa gcaacccaaa
     2401 gctatggggc ctaccccacc cagcccgggc agggctattc ccagcagagc agtcagccct
     2461 acggacagca gagttacagt ggttatagcc agtccacgga cacttcaggc tatggccaga
     2521 gcagctattc ttcttatggc cagagccaga acacaggcta tggaactcag tcaactcccc
     2581 agggatatgg ctcgactggc ggctatggca gtagccagag ctcccaatcg tcttacgggc
     2641 agcagtcctc ctatcctggc tatggccagc agccagctcc cagcagcacc tcgggaagtt
     2701 acggtagcag ttctcagagc agcagctatg ggcagcccca gagtgggagc tacagccagc
     2761 agcctagcta tggtggacag cagcaaagct atggacagca gcaaagctat aatccccctc
     2821 agggctatgg acagcagaac cagtacaaca gcagcagtgg tggtggaggt ggaggtggag
     2881 gtggaggtaa ctatggccaa gatcaatcct ccatgagtag tggtggtggc agtggtggcg
     2941 gttatggcaa tcaagaccag agtggtggag gtggcagcgg tggctatgga cagcaggacc
     3001 gtggaggccg cggcaggggt ggcagtggtg gcggcggcgg cggcggcggt ggtggttaca
     3061 accgcagcag tggtggctat gaacccagag gtcgtggagg tggccgtgga ggcagaggtg
     3121 gcatgggcgg aagtgaccgt ggtggcttca ataaatttgg tggccctcgg gaccaaggat
     3181 cacgtcatga ctccgaacag gataattcag acaacaacac catctttgtg caaggcctgg
     3241 gtgagaatgt tacaattgag tctgtggctg attacttcaa gcagattggt attattaaga
     3301 caaacaagaa aacgggacag cccatgatta atttgtacac agacagggaa actggcaagc
     3361 tgaagggaga ggcaacggtc tcttttgatg acccaccttc agctaaagca gctattgact
     3421 ggtttgatgg taaagaattc tccggaaatc ctatcaaggt ctcatttgct actcgccggg
     3481 cagactttaa tcggggtggt ggcaatggtc gtggaggccg agggcgagga ggacccatgg
     3541 gccgtggagg ctatggaggt ggtggcagtg gtggtggtgg ccgaggagga tttcccagtg
     3601 gaggtggtgg cggtggagga cagcagcgag ctggtgactg gaagtgtcct aatcccacct
     3661 gtgagaatat gaacttctct tggaggaatg aatgcaacca gtgtaaggcc cctaaaccag
     3721 atggcccagg agggggacca ggtggctctc acatgggggg taactacggg gatgatcgtc
     3781 gtggtggcag aggaggctat gatcgaggcg gctaccgggg ccgcggcggg gaccgtggag
     3841 gcttccgagg gggccggggt ggtggggaca gaggtggctt tggccctggc aagatggatt
     3901 ccaggggtga gcacagacag gatcgcaggg agaggccgta ttaatctaga gtcgacccgg
     3961 gcggccgctt ccctttagtg agggttaatg cttcgagcag acatgataag atacattgat
     4021 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     4081 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     4141 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
     4201 aacctctaca aatgtggtaa aatccgataa ggatcgatcc gggctggcgt aatagcgaag
     4261 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     4321 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4381 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4441 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4501 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4561 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4621 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4681 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4741 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     4801 tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg
     4861 aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     4921 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4981 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5041 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5101 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5161 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5221 ttggaggcct aggcttttgc aaaaagcttg attcttctga cacaacagtc tcgaacttaa
     5281 ggctagagcc accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     5341 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5401 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5461 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5521 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5581 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5641 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5701 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5761 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5821 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5881 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5941 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     6001 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6061 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     6121 gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt cattacatct
     6181 gtgtgttggt tttttgtgtg aatcgatagc gataaggatc cgcgtatggt gcactctcag
     6241 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     6301 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     6361 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     6421 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     6481 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6541 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6601 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6661 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6721 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6781 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6841 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6901 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6961 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     7021 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     7081 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     7141 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     7201 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     7261 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     7321 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     7381 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7441 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7501 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7561 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7621 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7681 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7741 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7801 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7861 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7921 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7981 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     8041 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     8101 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     8161 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     8221 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     8281 tgctcacatg gctcgacaga tcttcaatat tggccattag ccatattatt cattggttat
     8341 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     8401 tacatttata ttggctcatg tccaatatga ccgccatgtt gg
//
