LOCUS       sequence-423388-        8849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DCAF1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8849)
  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 8849)
  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: pBiT3.1-C_DCAF1:1-1507
FEATURES             Location/Qualifiers
     source          1..8849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2454..2465)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5912..6033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5923..5942)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(6148..6181)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6191..6238
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6270..7064
                     /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(6324..6343)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6934..6953
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(7222..7810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7302..7321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    7936..7984
                     /note="synthetic polyadenylation signal"
     CDS             complement(8008..8406)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      8403..8412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8437..8794)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8451..8586
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8603..8794
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     8751..8771
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggcga tcgcaacaag cttcgggaag
     1201 atctggagct agcggcgggg gatccggtgg gaattcgaca ccatgactac agtagtggta
     1261 catgtggact ccaaagctga gctcactacc ctgctggagc agtgggaaaa ggaacatggc
     1321 agtgggcagg acatggtacc tatccttacc aggatgtctc aattgattga aaaagaaact
     1381 gaagagtatc gtaaagggga tccagaccca tttgatgatc gacatcctgg tcgagctgat
     1441 ccagagtgta tgctgggcca cttgctgaga atactcttca agaatgatga tttcatgaat
     1501 gcactggtga atgcatatgt gatgacaagc cgagagcccc ctttaaacac tgcagcttgc
     1561 agactcctat tagacatcat gccagggctg gaaactgctg tcgtctttca agaaaaggag
     1621 ggaattgtcg agaatctttt caaatgggcc cgagaggccg atcaaccatt gaggacatat
     1681 tctactggac tgttaggagg tgctatggaa aatcaagaca ttgctgccaa ctatagagat
     1741 gaaaattcac agctggtggc aatagtgctt cgaagactga gggagctaca gctacaggaa
     1801 gtggctttgc ggcaggaaaa caagcgtccc agtccacgga agctctcttc tgaacccctt
     1861 ttgcctctgg atgaggaggc tgtggatatg gactatggtg acatggctgt agatgtagtg
     1921 gatggagacc aagaggaagc ttctggagac atggagatct cctttcatct tgattcaggc
     1981 cacaagacta gtagcagagt gaactcaaca accaaacctg aggatggagg attaaagaaa
     2041 aacaagtcag caaaacaggg tgacagagag aactttagga aagccaagca aaagttgggt
     2101 ttctcatctt ctgatccaga tcgcatgttt gttgagctgt ctaatagcag ttggtcagaa
     2161 atgtctccct gggtgattgg caccaattat accctttatc ctatgactcc tgctatcgag
     2221 cagcgactca ttctccaata tttgacccct ctaggagaat atcaggagct acttcccata
     2281 ttcatgcaac ttggatcacg ggagctgatg atgttctata ttgacctgaa gcaaactaat
     2341 gatgtcctgc ttacatttga ggcactaaag cacctagcat ctctcctgct ccataacaaa
     2401 tttgccacag aatttgttgc gcatggtgga gtacagaaat tactggaaat acctcgtcct
     2461 tctatggctg caaccggtgt gtctatgtgt ttgtattacc tatcctacaa tcaggatgcc
     2521 atggaaagag tttgcatgca tccccacaat gttctgtctg atgtggtgaa ctataccctg
     2581 tggttaatgg agtgttctca tgcttcagga tgctgccatg ctaccatgtt tttttcaatt
     2641 tgcttctcat ttcgggccgt cttggagctc tttgaccgct atgatggtct tcgtcgtctg
     2701 gtgaacttga tcagtacttt ggagattcta aatttggaag atcagggtgc acttctgagt
     2761 gatgatgaaa tatttgctag ccgccaaact gggaaacata cctgcatggc cttgcgcaaa
     2821 tactttgagg ctcacctggc cattaaattg gaacaagtga agcagtcact tcagaggact
     2881 gagggtggca ttcttgtcca cccacaaccc ccgtacaagg catgctcata tactcatgaa
     2941 cagattgtgg aaatgatgga atttttgata gaatatggcc cagcgcagct atattgggaa
     3001 ccagctgaag ttttcctcaa actttcttgt gtgcaactct tgttgcagct tatttctatt
     3061 gcctgcaatt ggaagaccta ttatgcaagg aatgacactg tgcgctttgc tttggatgtc
     3121 ctggctattc ttactgtggt gccaaaaatc cagctccagt tggcagaatc agtggacgtg
     3181 ttggatgagg ctggatctac agtctctact gtaggtatca gcattatttt gggagtggct
     3241 gagggtgagt tcttcatcca tgatgctgaa attcagaagt cagcacttca gattatcatc
     3301 aattgtgtgt gtggcccaga taaccgaata tccagtattg gtaaatttat ctctggtact
     3361 cctcggagaa agctgcctca gaaccctaaa agcagtgagc acaccctggc caagatgtgg
     3421 aatgtggttc agtccaacaa cggcatcaag gtgctcctgt ccttactgtc cattaagatg
     3481 cccatcacag atgcagacca aatccgggcc ctggcctgca aagccctagt gggcctgtct
     3541 cgcagtagca ctgtccggca gatcatcagt aaactgcccc ttttcagcag ctgccagatc
     3601 cagcagctga tgaaggagcc tgtgctgcag gacaagcgca gtgaccatgt caagttctgc
     3661 aagtatgctg ctgaactcat tgaacgggtg tcaggaaaac cacttctcat tggcactgat
     3721 gtttccctag cacgactgca gaaagcagat gttgttgccc agtcaaggat ctccttccct
     3781 gagaaagagc tgcttttgtt gatacgaaac catcttattt ctaaagggct tggagaaaca
     3841 gcaaccgtgc tgacaaaaga ggctgacctg cccatgactg ctgcctccca ttcttctgcc
     3901 tttaccccag tcactgctgc tgcttctcct gtctctctac cccgaacccc tcgtatcgct
     3961 aatggcattg caactcgtct gggcagccat gctgctgtgg gtgcctctgc gccttctgcc
     4021 cctactgctc atcctcagcc acggcccccc cagggtccgc tagctctgcc cggcccatct
     4081 tatgcaggca actccccttt gattggtaga atcagtttta tcagagagag gccatcaccc
     4141 tgcaatggca ggaaaatcag agtgttgcgg cagaagtcgg accatggtgc ctacagccaa
     4201 agcccagcca taaaaaaaca gctggacaga catcttcctt ccccacctac gctggacagt
     4261 ataatcacag agtatcttag agaacaacat gctcgctgca agaatccagt tgccacctgc
     4321 ccacctttct ccctctttac tcctcaccaa tgtcctgagc caaaacagag gcggcaagcg
     4381 ccaataaact ttacgtcaag gctaaaccgc agggcatcat ttccaaagta tggaggggtg
     4441 gatggcggat gctttgatag gcaccttatc tttagcagat tccgtcctat ttcagtgttc
     4501 cgggaagcca atgaagatga gagtggcttc acctgctgtg cattctcagc acgggagcgg
     4561 ttcctgatgc ttggcacctg cacagggcag ctgaagctct ataatgtgtt tagtggacag
     4621 gaggaggcca gctataactg tcacaactca gccatcacac atcttgaacc ttccagggat
     4681 gggtccttgc tgctgacatc tgctacttgg agccagcctt tgtctgcact ttggggaatg
     4741 aagtcagtat ttgatatgaa gcattccttc acagaagatc actatgttga gttcagtaag
     4801 cactcccagg atcgggtcat cggcacaaaa ggagacattg cccacattta tgatattcag
     4861 actggcaaca agctgttgac tctgtttaac ccagatcttg ccaacaacta caagaggaac
     4921 tgtgccacct ttaatcctac agatgatctt gtcttaaatg atggcgtcct ctgggatgtc
     4981 cgctctgcac aggccatcca caagtttgac aagttcaata tgaacatcag tggtgttttc
     5041 catccaaatg gactggaggt gatcattaat actgagattt gggaccttcg aacttttcat
     5101 cttttgcata ctgttcccgc tctggatcag tgtcgcgtgg tgttcaatca cacgggaaca
     5161 gtgatgtatg gagctatgtt gcaggcagat gatgaagatg acttaatgga agagaggatg
     5221 aaaagcccct ttgggtcatc cttccgaaca tttaatgcaa ctgactacaa acctatagca
     5281 accattgatg tgaaacggaa catctttgac ctgtgtacag acaccaaaga ctgctatctt
     5341 gctgtcattg agaatcaagg cagcatggat gccctgaaca tggacacagt atgcaggctg
     5401 tatgaagtgg gcaggcagcg tctggcagag gatgaggatg aagaggagga ccaggaagag
     5461 gaagaacagg aggaagaaga tgatgatgaa gatgatgatg acaccgatga tttagatgag
     5521 cttgacactg accagttgct ggaggcggag ttggaggagg acgacaataa tgagaacgca
     5581 ggggaagatg gggacaatga cttctctccc tctgatgagg agctagcaaa ccttctagag
     5641 gagggagagg acggggagga tgaagactct gatgcagatg aggaggtgga actgatcctg
     5701 ggggacactg acagctctga caactctgat ttggaagatg acatcatctt atctctgaat
     5761 gagggctcga gcggtgggag ctccggtgtg agcggctggc ggctgttcaa gaagattagc
     5821 taagtttaaa cggccgcgac tctagactgc aggcatgcaa gctgatttgg ctgctgccac
     5881 cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     5941 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     6001 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     6061 tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt
     6121 ggtaaaatct taattaagac ttctaatata acttcgtata gcatacatta tacgaagtta
     6181 tggttccact gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     6241 ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg cacgcaggtt
     6301 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     6361 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     6421 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     6481 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     6541 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     6601 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6661 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6721 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6781 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6841 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6901 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6961 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     7021 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa atgaccgacc
     7081 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7141 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7201 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7261 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7321 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7381 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7441 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7501 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7561 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7621 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7681 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7741 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7801 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7861 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7921 atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt
     7981 tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag agggcagcaa
     8041 ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg ctttgatccc
     8101 aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt
     8161 ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag cagcagtgcc
     8221 cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga caccagtgaa
     8281 gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag agatggggat
     8341 gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag acaaaggctt
     8401 ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta ggcctccaaa
     8461 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     8521 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     8581 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     8641 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     8701 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     8761 acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag ttcctctttc
     8821 agaggttatt tcaggccatg gtgctgcgc
//
