LOCUS       sequence-423298-        7758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DCAF15. C-terminal NanoLuc
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7758)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova YA, Baghaie AJ, Beldar S, Bolotokova A, Centrella
            PA, Chau I, Clark MA, Cuozzo JW, Dehghani-Tafti S, Disch JS, Dong A,
            Dumas A, Feng JA, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green SR,
            Guie MA, Guilinger JP, Harms N, Herasymenko O, Houliston S, 
            Hutchinson A, Kearnes S, Keefe AD, Kimani SW, Kramer T, Kutera M, 
            Kwak HA, Lento C, Li Y, Liu J, Loup J, Machado RAC, Mulhern CJ, 
            Perveen S, Righetto GL, Riley P, Shrestha S, Sigel EA, Silva M, 
            Sintchak MD, Slakman BL, Taylor RD, Thompson J, Torng W, 
            Underkoffler C, von Rechenberg M, Walsh RT, Watson I, Wilson DJ, 
            Wolf E, Yadav M, Yazdi AK, Zhang J, Zhang Y, Santhakumar V, Edwards 
            AM, Barsyte-Lovejoy D, Schapira M, Brown PJ, Halabelian L, 
            Arrowsmith CH
  TITLE     A Target Class Ligandability Evaluation of WD40 Repeat-Containing 
            Proteins.
  JOURNAL   J Med Chem. 2025 Jan 23;68(2):1092-1112. doi: 
            10.1021/acs.jmedchem.4c02010. Epub 2024 Nov 4.
  PUBMED    39495097
REFERENCE   2  (bases 1 to 7758)
  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"; doi: 
            "10.1021/acs.jmedchem.4c02010"; journalName: "J Med Chem"; date: 
            "2025-01-23- 23"; volume: "68"; issue: "2"; pages: "1092-1112"
COMMENT     Sequence Label: pNLF1-C_DCAF15:1-600
FEATURES             Location/Qualifiers
     source          1..7758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1079..1088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2933..3445
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3521..3539)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3576..3697
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3587..3606)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3641..3660
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3942..4299
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3942..4133
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3965..3985)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4150..4285
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4330..5367
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5346..5354
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5391..5439
                     /note="synthetic polyadenylation signal"
     promoter        5595..5699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5700..5768,5769..6560)
                     /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(5918..5937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6718..7306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6798..6817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7422..7705
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gcgatcgctt ccgaattcgc
     1081 caccatggcg cccagctcga aatcggagcg gaacagcggg gctgggagcg gcggcggcgg
     1141 ccccggggga gccggaggga agcgggcagc agggcggcgg cgggagcacg tcctcaagca
     1201 gctggagcgg gtcaagatca gcggacagct ctcccctcgc ctcttccgga agctgcctcc
     1261 ccgggtgtgc gtgtccctca agaacattgt ggatgaggac ttcctctatg caggacatat
     1321 cttcctgggc ttttccaaat gcggccgcta cgtcctctcc tacaccagca gcagtgggga
     1381 tgacgacttc tccttctaca tctaccatct gtactggtgg gagttcaacg ttcacagcaa
     1441 gctcaagctg gtccggcagg ttcggctatt ccaggacgag gagatctaca gcgacctgta
     1501 cctgaccgta tgcgagtggc ccagcgacgc ctccaaggtc atcgtcttcg gcttcaacac
     1561 ccgctcggcc aacgggatgc tcatgaacat gatgatgatg agtgacgaga accaccgtga
     1621 catctacgtc agcaccgtgg ccgtgccacc gccaggccgc tgtgctgctt gccaggatgc
     1681 cagccgagcc cacccaggag acccgaatgc acagtgccta cggcatggct tcatgctgca
     1741 caccaagtac caggtggtct accccttccc caccttccag cccgccttcc agctcaagaa
     1801 ggaccaggtg gtgctgctca acaccagcta ctccctggtg gcctgcgccg tctccgtcca
     1861 ctcggcaggt gacaggagtt tctgccaaat cctgtatgac cacagcacct gccccctggc
     1921 gcctgccagc ccccctgagc cccagagccc agagctgccc cctgccctcc ccagcttctg
     1981 ccctgaggcg gccccagccc gttcttctgg gtctcctgag ccctcgcccg ccattgccaa
     2041 agccaaggag tttgtggctg acatcttccg ccgggccaaa gaggccaagg gcggggtccc
     2101 tgaggaagcc cggcctgccc tgtgcccagg accctctggc agccgctgcc gtgcgcactc
     2161 tgagccccta gccctgtgtg gagagacggc accccgggac agcccccctg cctcggaggc
     2221 acctgcctcc gagcctggct atgtcaacta caccaagctg tactatgtgc tggagtccgg
     2281 agaggggacg gagccggagg atgagttgga ggacgacaag atctccctgc ccttcgtggt
     2341 gactgatctt cgtggccgca acctgcggcc catgcgggag cggactgctg tccagggcca
     2401 gtacctgaca gtggagcagc tcacactaga cttcgaatat gttatcaatg aggtcatccg
     2461 ccacgacgct acctggggcc atcagttctg ttctttcagc gactatgaca tcgtcattct
     2521 ggaggtctgc ccagaaacca accaggtcct catcaacatt ggcctgctgc tcctggcctt
     2581 cccgtccccc actgaggagg gccagctccg accaaagacc tatcacacca gcctcaaggt
     2641 ggcatgggac ctcaacacag ggatcttcga gacagtcagt gtaggcgacc tgactgaggt
     2701 caaagggcag accagcggca gtgtctggag ctcctaccgc aagagctgcg tggacatggt
     2761 catgaagtgg ctggtgccgg agagcagcgg ccgctacgtc aacaggatga ccaatgaggc
     2821 gctgcacaaa gggtgctccc tgaaggttct ggcggacagc gagcgatata cgtggatcgt
     2881 gctgtctaga tttgggccca attcctgcag gcgagctctc ggctcgagcg gcgtcttcac
     2941 actcgaagat ttcgttgggg actggcgaca gacagccggc tacaacctgg accaagtcct
     3001 tgaacaggga ggtgtgtcca gtttgtttca gaatctcggg gtgtccgtaa ctccgatcca
     3061 aaggattgtc ctgagcggtg aaaatgggct gaagatcgac atccatgtca tcatcccgta
     3121 tgaaggtctg agcggcgacc aaatgggcca gatcgaaaaa atttttaagg tggtgtaccc
     3181 tgtggatgat catcacttta aggtgatcct gcactatggc acactggtaa tcgacggggt
     3241 tacgccgaac atgatcgact atttcggacg gccgtatgaa ggcatcgccg tgttcgacgg
     3301 caaaaagatc actgtaacag ggaccctgtg gaacggcaac aaaattatcg acgagcgcct
     3361 gatcaacccc gacggctccc tgctgttccg agtaaccatc aacggagtga ccggctggcg
     3421 gctgtgcgaa cgcattctgg cgtaaggccg cgactctagg gcatgcaagc tgatccggct
     3481 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     3541 taaccccttg gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     3601 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3661 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3721 tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa
     3781 atgtggtaaa atcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     3841 tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg
     3901 cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag
     3961 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4021 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4081 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4141 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4201 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4261 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcgattc ttctgacact
     4321 agcgccacca tgaagaagcc cgaactcacc gctaccagcg ttgaaaaatt tctcatcgag
     4381 aagttcgaca gtgtgagcga cctgatgcag ttgtcggagg gcgaagagag ccgagccttc
     4441 agcttcgatg tcggcggacg cggctatgta ctgcgggtga atagctgcgc tgatggcttc
     4501 tacaaagacc gctacgtgta ccgccacttc gccagcgctg cactacccat ccccgaagtg
     4561 ttggacatcg gcgagttcag cgagagcctg acatactgca tcagtagacg cgcccaaggc
     4621 gttactctcc aagacctccc cgaaacagag ctgcctgctg tgttacagcc tgtcgccgaa
     4681 gctatggatg ctattgccgc cgccgacctc agtcaaacca gcggcttcgg cccattcggg
     4741 ccccaaggca tcggccagta cacaacctgg cgggatttca tttgcgccat tgctgatccc
     4801 catgtctacc actggcagac cgtgatggac gacaccgtgt ccgccagcgt agctcaagcc
     4861 ctggacgaac tgatgctgtg ggccgaagac tgtcccgagg tgcgccacct cgtccatgcc
     4921 gacttcggca gcaacaacgt cctgaccgac aacggccgca tcaccgccgt aatcgactgg
     4981 tccgaagcta tgttcgggga cagtcagtac gaggtggcca acatcttctt ctggcggccc
     5041 tggctggctt gcatggagca gcagactcgc tacttcgagc gccggcatcc cgagctggcc
     5101 ggcagccctc gtctgcgagc ctacatgctg cgcatcggcc tggatcagct ctaccagagc
     5161 ctcgtggacg gcaacttcga cgatgctgcc tgggctcaag gccgctgcga tgccatcgtc
     5221 cgcagcgggg ccggcaccgt cggtcgcaca caaatcgctc gccggagcgc agccgtatgg
     5281 accgacggct gcgtcgaggt gctggccgac agcggcaacc gccggcccag tacacgaccg
     5341 cgcgctaagg aggtaggtcg agtttagact ctagccatca cgatggccgc aataaaatat
     5401 ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc
     5461 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
     5521 agcaccgttt ctgcggactg gctttctacg taatggtttc ttagacgtca ggtggcactt
     5581 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5641 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5701 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5761 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5821 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5881 aagaacgttt tccaatgatg agcactttca aagttctgct atgtggcgcg gtattatccc
     5941 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6001 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6061 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaactatcg
     6121 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6181 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6241 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6301 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6361 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6421 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6481 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6541 cactgattaa gcattggtaa ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc
     6601 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6661 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6721 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6781 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6841 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6901 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6961 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7021 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7081 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7141 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7201 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7261 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat
     7321 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7381 gagattatca aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca
     7441 cgctggatgg cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta
     7501 agtgggggta atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta
     7561 tcaggcgcga ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc
     7621 gtgatcgcgc tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact
     7681 tactgtctgc cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg
     7741 cgtagaggat cgagatct
//
