LOCUS       sequence-170451-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  AUTHORS   Sun Z, Diaz Z, Fang X, Hart MP, Chesi A, Shorter J, Gitler AD
  TITLE     Molecular determinants and genetic modifiers of aggregation and 
            toxicity for the ALS disease protein FUS/TLS.
  JOURNAL   PLoS Biol. 2011 Apr;9(4):e1000614. doi: 
            10.1371/journal.pbio.1000614. Epub 2011 Apr 26.
  PUBMED    21541367
REFERENCE   2  (bases 1 to 7083)
  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: "PLoS 
            Biol."; date: "2011-04"; volume: "9(4):e1000614. doi"; pages: " 
            10.1371/journal.pbio.1000614. Epub 2011 Apr 26"
COMMENT     Sequence Label: pcDNA 3.2-FUS-1-526aa-V5
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    677..701
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1087..1098
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(2289..2313)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2339..2380
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2420..2439)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2464..2512
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2714..3142
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2801..2820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3011..3032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3151..3171)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3156..3485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3336..3471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3552..4346
                     /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(3606..3625)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4216..4235
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4522..4643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4559..4578)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4613..4632
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4692..4708)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4692..4708)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4705..4727)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4716..4732
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4740..4746,4747..4764,4765..4770))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4785..4806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4923..4940)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5094..5682)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5174..5193)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5853..6644,6645..6713))
                     /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     6476..6495
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6714..6818)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6893..6912)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag ctatcaacaa gtttgtacaa aaaagcaggc tccaccatgg cctcaaacga
      721 ttatacccaa caagcaaccc aaagctatgg ggcctacccc acccagcccg ggcagggcta
      781 ttcccagcag agcagtcagc cctacggaca gcagagttac agtggttata gccagtccac
      841 ggacacttca ggctatggcc agagcagcta ttcttcttat ggccagagcc agaacacagg
      901 ctatggaact cagtcaactc cccagggata tggctcgact ggcggctatg gcagtagcca
      961 gagctcccaa tcgtcttacg ggcagcagtc ctcctatcct ggctatggcc agcagccagc
     1021 tcccagcagc acctcgggaa gttacggtag cagttctcag agcagcagct atgggcagcc
     1081 ccagagtggg agctacagcc agcagcctag ctatggtgga cagcagcaaa gctatggaca
     1141 gcagcaaagc tataatcccc ctcagggcta tggacagcag aaccagtaca acagcagcag
     1201 tggtggtgga ggtggaggtg gaggtggagg taactatggc caagatcaat cctccatgag
     1261 tagtggtggt ggcagtggtg gcggttatgg caatcaagac cagagtggtg gaggtggcag
     1321 cggtggctat ggacagcagg accgtggagg ccgcggcagg ggtggcagtg gtggcggcgg
     1381 cggcggcggc ggtggtggtt acaaccgcag cagtggtggc tatgaaccca gaggtcgtgg
     1441 aggtggccgt ggaggcagag gtggcatggg cggaagtgac cgtggtggct tcaataaatt
     1501 tggtggccct cgggaccaag gatcacgtca tgactccgaa caggataatt cagacaacaa
     1561 caccatcttt gtgcaaggcc tgggtgagaa tgttacaatt gagtctgtgg ctgattactt
     1621 caagcagatt ggtattatta agacaaacaa gaaaacggga cagcccatga ttaatttgta
     1681 cacagacagg gaaactggca agctgaaggg agaggcaacg gtctcttttg atgacccacc
     1741 ttcagctaaa gcagctattg actggtttga tggtaaagaa ttctccggaa atcctatcaa
     1801 ggtctcattt gctactcgcc gggcagactt taatcggggt ggtggcaatg gtcgtggagg
     1861 ccgagggcga ggaggaccca tgggccgtgg aggctatgga ggtggtggca gtggtggtgg
     1921 tggccgagga ggatttccca gtggaggtgg tggcggtgga ggacagcagc gagctggtga
     1981 ctggaagtgt cctaatccca cctgtgagaa tatgaacttc tcttggagga atgaatgcaa
     2041 ccagtgtaag gcccctaaac cagatggccc aggaggggga ccaggtggct ctcacatggg
     2101 gggtaactac ggggatgatc gtcgtggtgg cagaggaggc tatgatcgag gcggctaccg
     2161 gggccgcggc ggggaccgtg gaggcttccg agggggccgg ggtggtgggg acagaggtgg
     2221 ctttggccct ggcaagatgg attccagggg tgagcacaga caggatcgca gggagaggcc
     2281 gtatttggac ccagctttct tgtacaaagt ggttgatcta gagggcccgc ggttcgaagg
     2341 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtt agtaatgagt
     2401 ttaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     2461 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     2521 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     2581 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     2641 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     2701 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2761 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2821 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2881 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2941 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3001 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3061 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3121 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3301 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3361 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3421 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3481 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3541 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3601 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3661 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3721 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3781 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3841 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3901 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3961 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4021 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4081 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4141 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4201 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4261 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4321 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     4381 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4441 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4501 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4561 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4621 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4681 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4741 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4801 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4861 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4921 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4981 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5041 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5101 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5161 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5221 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5281 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5341 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5401 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5461 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5521 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5581 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5641 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5701 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5761 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5821 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5881 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5941 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6001 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6061 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6121 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6181 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6241 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6301 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6361 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6421 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6481 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6541 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6601 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6661 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6721 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6781 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6841 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     6901 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     6961 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7021 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     7081 gtt
//
