LOCUS       sequence-494100-        8263 bp    DNA     circular SYN 01-DEC-2025
DEFINITION  Express DCAF5(1-477) in Insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8263)
  AUTHORS   Radko-Juettner S, Yue H, Myers JA, Carter RD, Robertson AN, Mittal 
            P, Zhu Z, Hansen BS, Donovan KA, Hunkeler M, Rosikiewicz W, Wu Z, 
            McReynolds MG, Roy Burman SS, Schmoker AM, Mageed N, Brown SA, 
            Mobley RJ, Partridge JF, Stewart EA, Pruett-Miller SM, Nabet B, Peng
            J, Gray NS, Fischer ES, Roberts CWM
  TITLE     Targeting DCAF5 suppresses SMARCB1-mutant cancer by stabilizing 
            SWI/SNF.
  JOURNAL   Nature. 2024 Apr;628(8007):442-449. doi: 10.1038/s41586-024-07250-1.
            Epub 2024 Mar 27.
  PUBMED    38538798
REFERENCE   2  (bases 1 to 8263)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41586-024-07250-1"; journalName: "Nature"; date: 
            "2024-04"; volume: "628"; issue: "8007"; pages: "442-449"
COMMENT     Sequence Label: pHoY108_pAC8_hsDCAF5_1-477
FEATURES             Location/Qualifiers
     source          1..8263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      57..645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     546..565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     799..816
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     1208..2035
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             1208..1332
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(1370..1975)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(2039..2072)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2262..2310
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2335..2354
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2388..3065)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Strongin et al., 2007)"
                     /label=DsRed-Monomer
                     /note="mammalian codon-optimized"
                     /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGV
                     VEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISH
                     ALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHS
                     GSQ"
     primer_bind     complement(2648..2667)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(3074..3183)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        3238..3329
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3254..3271
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             3347..3370
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3380..3424
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3443..3463
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_recomb     4943..6300
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(4959..6300)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
                     LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
                     PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
                     PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
                     APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
                     EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
                     PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
                     IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     rep_origin      6447..6902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6534..6553)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6744..6765
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6919..6938)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7038..7060
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7098..7116)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7184..7288
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7289..7357,7358..8149)
                     /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(7507..7526)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
       61 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
      121 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
      181 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
      241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
      301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
      361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
      421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
      481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
      541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
      601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
      661 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
      721 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
      781 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
      841 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gaacggctcc gcccactatt
      901 aatgaaatta aaaattccaa ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa
      961 tgcgtagaag gaaagaaaaa tgtcgtcgac atgctgaaca acaagattaa tatgcctccg
     1021 tgtataaaaa aaatattgaa cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac
     1081 aggaagaggt ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa
     1141 aaccgatgtt taatcaaggc tctgacgcat ttctacaacc acgactccaa gtgtgtgggt
     1201 gaagtcatgc atcttttaat caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa
     1261 atgaaaactg tcgacaagct ctgtccgttt gctggcaact gcaagggtct caatcctatt
     1321 tgtaattatt gaataataaa acaattataa atgtcaaatt tgttttttat taacgataca
     1381 aaccaaacgc aacaagaaca tttgtagtat tatctataat tgaaaacgcg tagttataat
     1441 cgctgaggta atatttaaaa tcattttcaa atgattcaca gttaatttgc gacaatataa
     1501 ttttattttc acataaacta gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt
     1561 catttttctc ctcataaaaa ttaacatagt tattatcgta tccatatatg tatctatcgt
     1621 atagagtaaa ttttttgttg tcataaatat atatgtcttt tttaatgggg tgtatagtac
     1681 cgctgcgcat agtttttctg taatttacaa cagtgctatt ttctggtagt tcttcggagt
     1741 gtgttgcttt aattattaaa tttatataat caatgaattt gggatcgtcg gttttgtaca
     1801 atatgttgcc ggcatagtac gcagcttctt ctagttcaat tacaccattt tttagcagca
     1861 ccggattaac ataactttcc aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc
     1921 ccttttctat actattgtct gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa
     1981 tgagacgcac aaactaatat cacaaactgg aaatgtctat caatatatag ttgctgatat
     2041 aacttcgtat agcatacatt atacgaagtt atctggttga aacgtacccg tagtggctat
     2101 ggcagggctt gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     2161 ttggggtggg gaaaaggaag aaacgcgggc gtattggtcc caatggggtc tcggtggggt
     2221 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     2281 cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct
     2341 ccttccgtgt ttcagttagc ctcccccatc tcccgctatt actcgagtca ctgggagccg
     2401 gagtggcggg cctcggcgtg ctcgtactgc tccaccacgg tgtagtcctc gttgtggttg
     2461 gtgatgtcca gcttggagtc cacgtagtgg ttgccgggca gctgcacggg cttcttggcc
     2521 ttgtacacgg tcttgaagtc gcaggtgtag tggccgccgt ccttcagctt cagggcgtgg
     2581 gagatctcgc ccttcagcac gccgtcctgg gggtacagct tctcggtgga gggctcccag
     2641 ccggcagtct tcttctgcat tacggggccg tcggcgggga agttcacgcc cttgaacttc
     2701 accttgtaga tgaaggtgcc gtcctgcagg gaggagtcct gctgcacctc caccacgccg
     2761 ccgtcctcga agttcatgga gcgctcccag gtgaagccct cggggaagga cagcttcatg
     2821 tagtcgggga tgtcggcggg gtgcttcacg taggccttgg agccgtactg gaactggggg
     2881 gacaggatgt cccaggcgaa gggcaggggg ccgcccttgg tcacctgcag cttggcggtc
     2941 tgggtgccct cgtagggctt gccctcgccc tcgccctcga tctcgaagta gtggccgttc
     3001 acggagccct ccatgcgcac cttgaactgc atgaactcct tgatgacgtc ctcggtgttg
     3061 tccatggtag agtgattgta aataaaatgt aatttacagt atagtatttt aattaatata
     3121 caaatgattt gataataatt cttatttaac tataatatat tgtgttgggt tgaattaaag
     3181 gtccgtatac tagtatcgat tcgcgaccta ctccggaata ttaatagagt tgctgatatc
     3241 atggagataa ttaaaatgat aaccatctcg caaataaata agtattttac tgttttcgta
     3301 acagttttgt aataaaaaaa cctataaata cggatctacc atggactgga gccacccgca
     3361 gttcgaaaaa tccgcggtcg gcctgaacga catcttcgag gctcagaaaa tcgaatggca
     3421 cgaaggtgga ggaggttctg gagagaacct ctacttccag ggaggcggcc gcatgaagag
     3481 gagggcggga ctgggaggaa gcatgagatc agtagtagga tttcttagtc agaggggatt
     3541 gcacggtgat ccattgctga cacaagattt ccagcgtagg cgtcttcgtg gatgccgtaa
     3601 tctttacaaa aaagacttgt tgggccactt cggctgcgta aatgcaattg agttcagcaa
     3661 taacggaggc cagtggctcg tgtcaggagg cgatgacaga agagttttgc tgtggcacat
     3721 ggagcaggcc atccatagtc gtgtaaaacc tattcaactt aaaggagaac atcattcaaa
     3781 tatcttttgt ttggcattca acagtggtaa tactaaagta ttctccggag gcaacgatga
     3841 acaggttata ctccatgacg tcgagtccag cgagacactg gatgtttttg cacatgagga
     3901 cgcagtttat ggtcttagcg taagccccgt aaatgataac atcttcgcgt cttcttcaga
     3961 cgacggcagg gtgcttatat gggatattag ggaatcaccc cacggagagc cattttgtct
     4021 cgccaattac cccagtgcgt tccacagcgt aatgttcaat cccgtagaac cgcgtctgct
     4081 tgcgacggcg aattcaaagg aaggagttgg actgtgggac atcagaaaac cacagagcag
     4141 tctcttgcgt tatggtggaa atctgagctt gcaatccgca atgtcagtgc gcttcaacag
     4201 taatggaact caacttctgg cgttgagacg taggttgccg cctgttctgt atgacataca
     4261 cagccgtctg cctgtctttc aatttgataa tcaaggctat tttaactcat gcacgatgaa
     4321 aagttgttgt ttcgcgggtg acagggacca atatatactc agcggctccg atgacttcaa
     4381 cttgtacatg tggaggatcc cagctgatcc ggaggcaggt ggtatcggcc gtgtagtcaa
     4441 tggtgctttc atggtactta agggccacag atctatagtg aatcaggttc gcttcaatcc
     4501 acatacttat atgatatgct catccggtgt agagaaaatt attaaaattt ggagtcccta
     4561 taagcagccg ggctgtaccg gtgaccttga tggcagaatc gaagacgatt ctagatgcct
     4621 ttacacgcac gaagaataca ttagtcttgt tcttaatagt ggatcaggtc tctcccacga
     4681 ttatgctaac caaagcgtac aggaagaccc gcgtatgatg gcatttttcg acagcctcgt
     4741 taggcgtgag atcgagggct ggagctcaga cagcgattca gatctttctg aaagcacgat
     4801 tcttcagctt cacgctggtg tgtccgaacg ttccggatat acggactcag agtctagtgc
     4861 gtcacttccc cgctccccac cgccgacagt tgatgagtca gcatgaggta cccccggacg
     4921 gccgcttaat taattgatcc gggttattag tacatttatt aagcgctaga ttctgtgcgt
     4981 tgttgattta cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta
     5041 gggtggtatg ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat
     5101 attaaatcct caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt
     5161 tccgaaccga tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa
     5221 tcttgtagca gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt
     5281 tcgacgtcgt tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac
     5341 aattgactcg acgtaaacac gttaaataaa gcttggacat atttaacatc gggcgtgtta
     5401 gctttattag gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa
     5461 gacgattttg ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc
     5521 aaatttgtag ttgagctttt tggaattatt tctgattgcg ggcgtttttg ggcgggtttc
     5581 aatctaactg tgcccgattt taattcagac aacacgttag aaagcgatgg tgcaggcggt
     5641 ggtaacattt cagacggcaa atctactaat ggcggcggtg gtggagctga tgataaatct
     5701 accatcggtg gaggcgcagg cggggctggc ggcggaggcg gaggcggagg tggtggcggt
     5761 gatgcagacg gcggtttagg ctcaaatgtc tctttaggca acacagtcgg cacctcaact
     5821 attgtactgg tttcgggcgc cgtttttggt ttgaccggtc tgagacgagt gcgatttttt
     5881 tcgtttctaa tagcttccaa caattgttgt ctgtcgtcta aaggtgcagc gggttgaggt
     5941 tccgtcggca ttggtggagc gggcggcaat tcagacatcg atggtggtgg tggtggtgga
     6001 ggcgctggaa tgttaggcac gggagaaggt ggtggcggcg gtgccgccgg tataatttgt
     6061 tctggtttag tttgttcgcg cacgattgtg ggcaccggcg caggcgccgc tggctgcaca
     6121 acggaaggtc gtctgcttcg aggcagcgct tggggtggtg gcaattcaat attataattg
     6181 gaatacaaat cgtaaaaatc tgctataagc attgtaattt cgctatcgtt taccgtgccg
     6241 atatttaaca accgctcaat gtaagcaatt gtattgtaaa gagattgtct caagctcgga
     6301 tcgatcccgc acgccgataa caagcctttt catttttact acagcattgt agtggcgaga
     6361 cacttcgctg tcgtcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     6421 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     6481 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     6541 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6601 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     6661 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     6721 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     6781 tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     6841 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     6901 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     6961 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     7021 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     7081 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     7141 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     7201 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     7261 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     7321 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7381 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7441 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     7501 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     7561 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7621 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7681 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     7741 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7801 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7861 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7921 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7981 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     8041 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     8101 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     8161 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     8221 ggtgaagatc ctttttgata atctcatgac caaaatccct taa
//
