LOCUS       sequence-423297-        7707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DCAF15. N-terminal NanoLuc
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7707)
  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 7707)
  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-N_DCAF15:1-600
FEATURES             Location/Qualifiers
     source          1..7707
                     /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"
     CDS             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3475..3493)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3525..3646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3536..3555)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3590..3609
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3891..4248
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3891..4082
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3914..3934)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4099..4234
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4279..5316
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5295..5303
                     /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    5340..5388
                     /note="synthetic polyadenylation signal"
     promoter        5544..5648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5649..5717,5718..6509)
                     /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(5867..5886)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6667..7255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6747..6766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7371..7654
                     /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 ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgcttc cgaattcatg gcgcccagct
     1621 cgaaatcgga gcggaacagc ggggctggga gcggcggcgg cggccccggg ggagccggag
     1681 ggaagcgggc agcagggcgg cggcgggagc acgtcctcaa gcagctggag cgggtcaaga
     1741 tcagcggaca gctctcccct cgcctcttcc ggaagctgcc tccccgggtg tgcgtgtccc
     1801 tcaagaacat tgtggatgag gacttcctct atgcaggaca tatcttcctg ggcttttcca
     1861 aatgcggccg ctacgtcctc tcctacacca gcagcagtgg ggatgacgac ttctccttct
     1921 acatctacca tctgtactgg tgggagttca acgttcacag caagctcaag ctggtccggc
     1981 aggttcggct attccaggac gaggagatct acagcgacct gtacctgacc gtatgcgagt
     2041 ggcccagcga cgcctccaag gtcatcgtct tcggcttcaa cacccgctcg gccaacggga
     2101 tgctcatgaa catgatgatg atgagtgacg agaaccaccg tgacatctac gtcagcaccg
     2161 tggccgtgcc accgccaggc cgctgtgctg cttgccagga tgccagccga gcccacccag
     2221 gagacccgaa tgcacagtgc ctacggcatg gcttcatgct gcacaccaag taccaggtgg
     2281 tctacccctt ccccaccttc cagcccgcct tccagctcaa gaaggaccag gtggtgctgc
     2341 tcaacaccag ctactccctg gtggcctgcg ccgtctccgt ccactcggca ggtgacagga
     2401 gtttctgcca aatcctgtat gaccacagca cctgccccct ggcgcctgcc agcccccctg
     2461 agccccagag cccagagctg ccccctgccc tccccagctt ctgccctgag gcggccccag
     2521 cccgttcttc tgggtctcct gagccctcgc ccgccattgc caaagccaag gagtttgtgg
     2581 ctgacatctt ccgccgggcc aaagaggcca agggcggggt ccctgaggaa gcccggcctg
     2641 ccctgtgccc aggaccctct ggcagccgct gccgtgcgca ctctgagccc ctagccctgt
     2701 gtggagagac ggcaccccgg gacagccccc ctgcctcgga ggcacctgcc tccgagcctg
     2761 gctatgtcaa ctacaccaag ctgtactatg tgctggagtc cggagagggg acggagccgg
     2821 aggatgagtt ggaggacgac aagatctccc tgcccttcgt ggtgactgat cttcgtggcc
     2881 gcaacctgcg gcccatgcgg gagcggactg ctgtccaggg ccagtacctg acagtggagc
     2941 agctcacact agacttcgaa tatgttatca atgaggtcat ccgccacgac gctacctggg
     3001 gccatcagtt ctgttctttc agcgactatg acatcgtcat tctggaggtc tgcccagaaa
     3061 ccaaccaggt cctcatcaac attggcctgc tgctcctggc cttcccgtcc cccactgagg
     3121 agggccagct ccgaccaaag acctatcaca ccagcctcaa ggtggcatgg gacctcaaca
     3181 cagggatctt cgagacagtc agtgtaggcg acctgactga ggtcaaaggg cagaccagcg
     3241 gcagtgtctg gagctcctac cgcaagagct gcgtggacat ggtcatgaag tggctggtgc
     3301 cggagagcag cggccgctac gtcaacagga tgaccaatga ggcgctgcac aaagggtgct
     3361 ccctgaaggt tctggcggac agcgagcgat atacgtggat cgtgctgtaa taatctagat
     3421 ttgggcccaa ttcctgcagg attttgcggc cgcttgctga gttggctgct gccaccgctg
     3481 agcaataact agcataaccc cttggccgct tcgagcagac atgataagat acattgatga
     3541 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3601 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3661 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     3721 cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat
     3781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca
     3841 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     3901 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3961 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4021 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4081 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4141 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4201 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgattct
     4261 tctgacacta gcgccaccat gaagaagccc gaactcaccg ctaccagcgt tgaaaaattt
     4321 ctcatcgaga agttcgacag tgtgagcgac ctgatgcagt tgtcggaggg cgaagagagc
     4381 cgagccttca gcttcgatgt cggcggacgc ggctatgtac tgcgggtgaa tagctgcgct
     4441 gatggcttct acaaagaccg ctacgtgtac cgccacttcg ccagcgctgc actacccatc
     4501 cccgaagtgt tggacatcgg cgagttcagc gagagcctga catactgcat cagtagacgc
     4561 gcccaaggcg ttactctcca agacctcccc gaaacagagc tgcctgctgt gttacagcct
     4621 gtcgccgaag ctatggatgc tattgccgcc gccgacctca gtcaaaccag cggcttcggc
     4681 ccattcgggc cccaaggcat cggccagtac acaacctggc gggatttcat ttgcgccatt
     4741 gctgatcccc atgtctacca ctggcagacc gtgatggacg acaccgtgtc cgccagcgta
     4801 gctcaagccc tggacgaact gatgctgtgg gccgaagact gtcccgaggt gcgccacctc
     4861 gtccatgccg acttcggcag caacaacgtc ctgaccgaca acggccgcat caccgccgta
     4921 atcgactggt ccgaagctat gttcggggac agtcagtacg aggtggccaa catcttcttc
     4981 tggcggccct ggctggcttg catggagcag cagactcgct acttcgagcg ccggcatccc
     5041 gagctggccg gcagccctcg tctgcgagcc tacatgctgc gcatcggcct ggatcagctc
     5101 taccagagcc tcgtggacgg caacttcgac gatgctgcct gggctcaagg ccgctgcgat
     5161 gccatcgtcc gcagcggggc cggcaccgtc ggtcgcacac aaatcgctcg ccggagcgca
     5221 gccgtatgga ccgacggctg cgtcgaggtg ctggccgaca gcggcaaccg ccggcccagt
     5281 acacgaccgc gcgctaagga ggtaggtcga gtttagactc tagccatcac gatggccgca
     5341 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga
     5401 taaggatcct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
     5461 tccggggtca gcaccgtttc tgcggactgg ctttctacgt aatggtttct tagacgtcag
     5521 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5581 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5641 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5701 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5761 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5821 ttcgccccga agaacgtttt ccaatgatga gcactttcaa agttctgcta tgtggcgcgg
     5881 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5941 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6001 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6061 caactatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6121 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6181 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6241 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6301 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6361 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6421 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6481 taggtgcctc actgattaag cattggtaat tcgaaatgac cgaccaagcg acgcccaacc
     6541 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6601 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6661 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6721 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6781 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6841 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6901 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6961 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7021 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7081 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7141 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7201 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
     7261 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7321 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt ccggaaatac
     7381 aggaacgcac gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc
     7441 agtggattaa gtgggggtaa tgtggcctgt accctctggt tgcataggta ttcatacggt
     7501 taaaatttat caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca tttttacctg
     7561 tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag ggattatggt
     7621 aaatccactt actgtctgcc ctcgtagcca tcgagataaa ccgcagtact ccggccacga
     7681 tgcgtccggc gtagaggatc gagatct
//
