LOCUS       sequence-362347-        8442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses mVenus and mCherry tagged to mutant of the FUS (R521C) 
            protein 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-R521C
FEATURES             Location/Qualifiers
     source          1..8442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..795
                     /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(55..74)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     665..684
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    859..907
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(948..967)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1067..1089
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1127..1145)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1213..1317
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1318..1386,1387..2178)
                     /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(1536..1555)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2349..2937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2838..2857
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3148..3527
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3528..3739
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3690..3710
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3736..3760
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          3900..4032
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4015..4034
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        4077..4095
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4115..4141
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(4142..4144,4145..4147,4148..4660)
                     /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(4187..4208)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4448..4467)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4795..4816
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4956..4974)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5224..5243
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5893..5904
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    7156..7277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7167..7186)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7221..7240
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      7463..7918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7550..7569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7760..7781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8035..8392
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        8035..8226
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(8058..8078)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      8243..8378
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
       61 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
      121 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
      181 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
      241 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
      301 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
      361 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
      421 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
      481 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
      541 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
      601 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
      661 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
      721 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
      781 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
      841 tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
      901 ttgtgtgaat cgatagcgat aaggatccgc gtatggtgca ctctcagtac aatctgctct
      961 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     1021 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     1081 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     1141 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1201 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1261 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1321 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1381 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1441 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1501 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1561 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1621 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1681 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1741 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1801 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1861 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1921 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1981 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2041 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2101 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2161 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2221 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2281 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2341 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2401 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2461 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2521 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2581 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2641 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2701 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2761 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2821 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2881 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2941 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct
     3001 cgacagatct tcaatattgg ccattagcca tattattcat tggttatata gcataaatca
     3061 atattggcta ttggccattg catacgttgt atctatatca taatatgtac atttatattg
     3121 gctcatgtcc aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat
     3181 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     3241 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     3301 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     3361 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg
     3421 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact
     3481 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     3541 ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     3601 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     3661 gtaacaactg cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     3721 ggtctatata agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt
     3781 agtttatcac agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt
     3841 aagctgcagt gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg
     3901 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     3961 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     4021 ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat
     4081 acgactcact ataggctagc ctcgagccac catgtaccca tacgacgtcc cagactacgc
     4141 tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     4201 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     4261 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
     4321 cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
     4381 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     4441 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     4501 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     4561 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     4621 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     4681 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     4741 ctacctgagc taccagtcca aactgagcaa agaccccaac gagaagcgcg atcacatggt
     4801 cctgctggag ttcgtgaccg ccgcctccgg agataacatg gccatcatca aggagttcat
     4861 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     4921 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     4981 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     5041 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     5101 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     5161 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     5221 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     5281 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     5341 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     5401 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     5461 acagtacgaa cgcgccgagg gccgccactc caccggcggc atgggctccc ttatggcctc
     5521 aaacgattat acccaacaag caacccaaag ctatggggcc taccccaccc agcccgggca
     5581 gggctattcc cagcagagca gtcagcccta cggacagcag agttacagtg gttatagcca
     5641 gtccacggac acttcaggct atggccagag cagctattct tcttatggcc agagccagaa
     5701 cacaggctat ggaactcagt caactcccca gggatatggc tcgactggcg gctatggcag
     5761 tagccagagc tcccaatcgt cttacgggca gcagtcctcc tatcctggct atggccagca
     5821 gccagctccc agcagcacct cgggaagtta cggtagcagt tctcagagca gcagctatgg
     5881 gcagccccag agtgggagct acagccagca gcctagctat ggtggacagc agcaaagcta
     5941 tggacagcag caaagctata atccccctca gggctatgga cagcagaacc agtacaacag
     6001 cagcagtggt ggtggaggtg gaggtggagg tggaggtaac tatggccaag atcaatcctc
     6061 catgagtagt ggtggtggca gtggtggcgg ttatggcaat caagaccaga gtggtggagg
     6121 tggcagcggt ggctatggac agcaggaccg tggaggccgc ggcaggggtg gcagtggtgg
     6181 cggcggcggc ggcggcggtg gtggttacaa ccgcagcagt ggtggctatg aacccagagg
     6241 tcgtggaggt ggccgtggag gcagaggtgg catgggcgga agtgaccgtg gtggcttcaa
     6301 taaatttggt ggccctcggg accaaggatc acgtcatgac tccgaacagg ataattcaga
     6361 caacaacacc atctttgtgc aaggcctggg tgagaatgtt acaattgagt ctgtggctga
     6421 ttacttcaag cagattggta ttattaagac aaacaagaaa acgggacagc ccatgattaa
     6481 tttgtacaca gacagggaaa ctggcaagct gaagggagag gcaacggtct cttttgatga
     6541 cccaccttca gctaaagcag ctattgactg gtttgatggt aaagaattct ccggaaatcc
     6601 tatcaaggtc tcatttgcta ctcgccgggc agactttaat cggggtggtg gcaatggtcg
     6661 tggaggccga gggcgaggag gacccatggg ccgtggaggc tatggaggtg gtggcagtgg
     6721 tggtggtggc cgaggaggat ttcccagtgg aggtggtggc ggtggaggac agcagcgagc
     6781 tggtgactgg aagtgtccta atcccacctg tgagaatatg aacttctctt ggaggaatga
     6841 atgcaaccag tgtaaggccc ctaaaccaga tggcccagga gggggaccag gtggctctca
     6901 catggggggt aactacgggg atgatcgtcg tggtggcaga ggaggctatg atcgaggcgg
     6961 ctaccggggc cgcggcgggg accgtggagg cttccgaggg ggccggggtg gtggggacag
     7021 aggtggcttt ggccctggca agatggattc caggggtgag cacagacagg attgcaggga
     7081 gaggccgtat taatctagag tcgacccggg cggccgcttc cctttagtga gggttaatgc
     7141 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     7201 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     7261 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     7321 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atccgataag
     7381 gatcgatccg ggctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     7441 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     7501 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     7561 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     7621 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     7681 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     7741 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     7801 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     7861 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     7921 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg
     7981 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     8041 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     8101 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     8161 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     8221 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     8281 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     8341 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttga
     8401 ttcttctgac acaacagtct cgaacttaag gctagagcca cc
//
