LOCUS       sequence-423385-        8805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DCAF1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8805)
  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 8805)
  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-N_DCAF1:1-1507
FEATURES             Location/Qualifiers
     source          1..8805
                     /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(2461..2472)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5868..5989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5879..5898)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(6104..6137)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6147..6194
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6226..7020
                     /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(6280..6299)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6890..6909
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(7178..7766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7258..7277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    7892..7940
                     /note="synthetic polyadenylation signal"
     CDS             complement(7964..8362)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      8359..8368
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8393..8750)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8407..8542
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8559..8750
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     8707..8727
                     /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 ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggta tgactacagt
     1261 agtggtacat gtggactcca aagctgagct cactaccctg ctggagcagt gggaaaagga
     1321 acatggcagt gggcaggaca tggtacctat ccttaccagg atgtctcaat tgattgaaaa
     1381 agaaactgaa gagtatcgta aaggggatcc agacccattt gatgatcgac atcctggtcg
     1441 agctgatcca gagtgtatgc tgggccactt gctgagaata ctcttcaaga atgatgattt
     1501 catgaatgca ctggtgaatg catatgtgat gacaagccga gagccccctt taaacactgc
     1561 agcttgcaga ctcctattag acatcatgcc agggctggaa actgctgtcg tctttcaaga
     1621 aaaggaggga attgtcgaga atcttttcaa atgggcccga gaggccgatc aaccattgag
     1681 gacatattct actggactgt taggaggtgc tatggaaaat caagacattg ctgccaacta
     1741 tagagatgaa aattcacagc tggtggcaat agtgcttcga agactgaggg agctacagct
     1801 acaggaagtg gctttgcggc aggaaaacaa gcgtcccagt ccacggaagc tctcttctga
     1861 accccttttg cctctggatg aggaggctgt ggatatggac tatggtgaca tggctgtaga
     1921 tgtagtggat ggagaccaag aggaagcttc tggagacatg gagatctcct ttcatcttga
     1981 ttcaggccac aagactagta gcagagtgaa ctcaacaacc aaacctgagg atggaggatt
     2041 aaagaaaaac aagtcagcaa aacagggtga cagagagaac tttaggaaag ccaagcaaaa
     2101 gttgggtttc tcatcttctg atccagatcg catgtttgtt gagctgtcta atagcagttg
     2161 gtcagaaatg tctccctggg tgattggcac caattatacc ctttatccta tgactcctgc
     2221 tatcgagcag cgactcattc tccaatattt gacccctcta ggagaatatc aggagctact
     2281 tcccatattc atgcaacttg gatcacggga gctgatgatg ttctatattg acctgaagca
     2341 aactaatgat gtcctgctta catttgaggc actaaagcac ctagcatctc tcctgctcca
     2401 taacaaattt gccacagaat ttgttgcgca tggtggagta cagaaattac tggaaatacc
     2461 tcgtccttct atggctgcaa ccggtgtgtc tatgtgtttg tattacctat cctacaatca
     2521 ggatgccatg gaaagagttt gcatgcatcc ccacaatgtt ctgtctgatg tggtgaacta
     2581 taccctgtgg ttaatggagt gttctcatgc ttcaggatgc tgccatgcta ccatgttttt
     2641 ttcaatttgc ttctcatttc gggccgtctt ggagctcttt gaccgctatg atggtcttcg
     2701 tcgtctggtg aacttgatca gtactttgga gattctaaat ttggaagatc agggtgcact
     2761 tctgagtgat gatgaaatat ttgctagccg ccaaactggg aaacatacct gcatggcctt
     2821 gcgcaaatac tttgaggctc acctggccat taaattggaa caagtgaagc agtcacttca
     2881 gaggactgag ggtggcattc ttgtccaccc acaacccccg tacaaggcat gctcatatac
     2941 tcatgaacag attgtggaaa tgatggaatt tttgatagaa tatggcccag cgcagctata
     3001 ttgggaacca gctgaagttt tcctcaaact ttcttgtgtg caactcttgt tgcagcttat
     3061 ttctattgcc tgcaattgga agacctatta tgcaaggaat gacactgtgc gctttgcttt
     3121 ggatgtcctg gctattctta ctgtggtgcc aaaaatccag ctccagttgg cagaatcagt
     3181 ggacgtgttg gatgaggctg gatctacagt ctctactgta ggtatcagca ttattttggg
     3241 agtggctgag ggtgagttct tcatccatga tgctgaaatt cagaagtcag cacttcagat
     3301 tatcatcaat tgtgtgtgtg gcccagataa ccgaatatcc agtattggta aatttatctc
     3361 tggtactcct cggagaaagc tgcctcagaa ccctaaaagc agtgagcaca ccctggccaa
     3421 gatgtggaat gtggttcagt ccaacaacgg catcaaggtg ctcctgtcct tactgtccat
     3481 taagatgccc atcacagatg cagaccaaat ccgggccctg gcctgcaaag ccctagtggg
     3541 cctgtctcgc agtagcactg tccggcagat catcagtaaa ctgccccttt tcagcagctg
     3601 ccagatccag cagctgatga aggagcctgt gctgcaggac aagcgcagtg accatgtcaa
     3661 gttctgcaag tatgctgctg aactcattga acgggtgtca ggaaaaccac ttctcattgg
     3721 cactgatgtt tccctagcac gactgcagaa agcagatgtt gttgcccagt caaggatctc
     3781 cttccctgag aaagagctgc ttttgttgat acgaaaccat cttatttcta aagggcttgg
     3841 agaaacagca accgtgctga caaaagaggc tgacctgccc atgactgctg cctcccattc
     3901 ttctgccttt accccagtca ctgctgctgc ttctcctgtc tctctacccc gaacccctcg
     3961 tatcgctaat ggcattgcaa ctcgtctggg cagccatgct gctgtgggtg cctctgcgcc
     4021 ttctgcccct actgctcatc ctcagccacg gcccccccag ggtccgctag ctctgcccgg
     4081 cccatcttat gcaggcaact cccctttgat tggtagaatc agttttatca gagagaggcc
     4141 atcaccctgc aatggcagga aaatcagagt gttgcggcag aagtcggacc atggtgccta
     4201 cagccaaagc ccagccataa aaaaacagct ggacagacat cttccttccc cacctacgct
     4261 ggacagtata atcacagagt atcttagaga acaacatgct cgctgcaaga atccagttgc
     4321 cacctgccca cctttctccc tctttactcc tcaccaatgt cctgagccaa aacagaggcg
     4381 gcaagcgcca ataaacttta cgtcaaggct aaaccgcagg gcatcatttc caaagtatgg
     4441 aggggtggat ggcggatgct ttgataggca ccttatcttt agcagattcc gtcctatttc
     4501 agtgttccgg gaagccaatg aagatgagag tggcttcacc tgctgtgcat tctcagcacg
     4561 ggagcggttc ctgatgcttg gcacctgcac agggcagctg aagctctata atgtgtttag
     4621 tggacaggag gaggccagct ataactgtca caactcagcc atcacacatc ttgaaccttc
     4681 cagggatggg tccttgctgc tgacatctgc tacttggagc cagcctttgt ctgcactttg
     4741 gggaatgaag tcagtatttg atatgaagca ttccttcaca gaagatcact atgttgagtt
     4801 cagtaagcac tcccaggatc gggtcatcgg cacaaaagga gacattgccc acatttatga
     4861 tattcagact ggcaacaagc tgttgactct gtttaaccca gatcttgcca acaactacaa
     4921 gaggaactgt gccaccttta atcctacaga tgatcttgtc ttaaatgatg gcgtcctctg
     4981 ggatgtccgc tctgcacagg ccatccacaa gtttgacaag ttcaatatga acatcagtgg
     5041 tgttttccat ccaaatggac tggaggtgat cattaatact gagatttggg accttcgaac
     5101 ttttcatctt ttgcatactg ttcccgctct ggatcagtgt cgcgtggtgt tcaatcacac
     5161 gggaacagtg atgtatggag ctatgttgca ggcagatgat gaagatgact taatggaaga
     5221 gaggatgaaa agcccctttg ggtcatcctt ccgaacattt aatgcaactg actacaaacc
     5281 tatagcaacc attgatgtga aacggaacat ctttgacctg tgtacagaca ccaaagactg
     5341 ctatcttgct gtcattgaga atcaaggcag catggatgcc ctgaacatgg acacagtatg
     5401 caggctgtat gaagtgggca ggcagcgtct ggcagaggat gaggatgaag aggaggacca
     5461 ggaagaggaa gaacaggagg aagaagatga tgatgaagat gatgatgaca ccgatgattt
     5521 agatgagctt gacactgacc agttgctgga ggcggagttg gaggaggacg acaataatga
     5581 gaacgcaggg gaagatgggg acaatgactt ctctccctct gatgaggagc tagcaaacct
     5641 tctagaggag ggagaggacg gggaggatga agactctgat gcagatgagg aggtggaact
     5701 gatcctgggg gacactgaca gctctgacaa ctctgatttg gaagatgaca tcatcttatc
     5761 tctgaatgag taatctagag gtcaagcttc gggggtttaa acctgcaggc atgcaagctg
     5821 atttggctgc tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga
     5881 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     5941 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     6001 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     6061 aaacctctac aaatgtggta aaatcttaat taagacttct aatataactt cgtatagcat
     6121 acattatacg aagttatggt tccactgttg acaattaatc atcggcatag tatatcggca
     6181 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat
     6241 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6301 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6361 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     6421 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6481 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6541 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6601 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6661 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6721 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     6781 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6841 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6901 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6961 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     7021 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     7081 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7141 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7201 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7261 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7321 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7381 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7441 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7501 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7561 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7621 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7681 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7741 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     7801 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7861 tcacctagat ccttttatag tccggaatcg tcacacaaaa aaccaacaca cagatgtaat
     7921 gaaaataaag atattttatt gcggccatcg tgatggctag agtttagccc tcccacacat
     7981 aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca
     8041 ctatggcttt gatcccagga tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc
     8101 tcaagatgcc cctgttctca tttccgatcg cgacgataca agtcaggttg ccagctgccg
     8161 cagcagcagc agtgcccagc accacgagtt ctgcacaagg tcccccagta aaatgatata
     8221 cattgacacc agtgaagatg cggccgtcgc tagagagagc tgcgctggcg acgctgtagt
     8281 cttcagagat ggggatgctg ttgattgtag ccgttgctct ttcaatgagg gtggattctt
     8341 cttgagacaa aggcttggcc atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa
     8401 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     8461 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     8521 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     8581 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     8641 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     8701 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag aaggtaccta
     8761 accaagttcc tctttcagag gttatttcag gccatggtgc tgcgc
//
