LOCUS       sequence-423422-        7154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DCAF5. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7154)
  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 7154)
  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_DCAF5:1-942
FEATURES             Location/Qualifiers
     source          1..7154
                     /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"
     polyA_signal    4217..4338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4228..4247)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4453..4486)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4496..4543
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4575..5369
                     /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(4629..4648)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5239..5258
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5527..6115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5607..5626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6241..6289
                     /note="synthetic polyadenylation signal"
     CDS             complement(6313..6711)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6708..6717
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6742..7099)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6756..6891
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6908..7099
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7056..7076
                     /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 ccatgaagag gagagctggc
     1261 ctggggggca gcatgaggtc agtggtgggc ttcttgtccc agcggggctt gcatggggac
     1321 cccctgctca ctcaggactt tcagaggaga cgcctgcggg gctgcagaaa cctctacaag
     1381 aaggacctcc tcggccactt cggctgtgtc aatgccattg aattctccaa caatggaggc
     1441 cagtggctgg tctcaggagg agatgaccgc cgggttctgc tatggcacat ggaacaagcc
     1501 atccactcca gggtcaagcc catacagctg aaaggagagc accattccaa cattttttgc
     1561 ctggctttca acagtgggaa cactaaagtg ttctctggag gcaatgatga gcaagttatc
     1621 ctccatgatg ttgaaagcag tgagacattg gacgtgtttg ctcatgaaga tgcagtatat
     1681 ggcttgtctg tgagcccagt gaatgacaac atttttgcca gttcctcaga tgatggccgg
     1741 gttctcattt gggacattcg ggaatccccc catggagagc ccttctgcct ggcaaactat
     1801 ccatcagcct ttcatagtgt catgtttaac cctgtggagc ccaggttgtt ggccacagcc
     1861 aattcaaagg aaggagtggg actctgggac attcgaaaac ctcagagttc tctcctgcgc
     1921 tatggtggaa acctgtccct ccaaagtgcc atgagtgtac gattcaacag caacgggacc
     1981 cagctcctgg ccctgaggcg acgcctgccc cctgtgctct atgacatcca ttcccgcctg
     2041 cctgtgtttc agtttgacaa tcagggttac ttcaactcat gcaccatgaa aagctgctgt
     2101 tttgcaggag atcgtgacca gtatatcctt tcgggctctg atgacttcaa cctgtacatg
     2161 tggagaattc ctgcagatcc agaagcaggt ggcattggta gggtggtcaa cggagccttc
     2221 atggtgctga aagggcatcg atctattgtt aaccaagtcc gatttaatcc ccacacctac
     2281 atgatctgct cttctggtgt agaaaagatt atcaagatct ggagcccata caagcagcca
     2341 ggatgtactg gagacctcga cggtcggatt gaggacgatt cccgctgcct ctatacccat
     2401 gaagagtaca tcagccttgt gctgaacagt gggagtggcc tgtcgcatga ctacgccaac
     2461 cagtcggtcc aggaagaccc ccggatgatg gccttctttg actcactggt acgccgagag
     2521 atcgagggct ggagctctga ctcagacagt gacctcagtg agagtactat cctccaactg
     2581 cacgctgggg tcagcgagcg ctcaggctac actgactcag agtcttcggc ctcattgcct
     2641 cgctccccgc ctcccacagt agatgagtct gccgacaacg ccttccacct ggggcccctg
     2701 cgggtcacca ccacaaacac agtagcctca actccaccaa cacccacgtg tgaggatgca
     2761 gcctctcgcc agcagcgtct gtctgctctg cggcgctacc aagacaaacg cctcctggcc
     2821 ctttccaatg agtccgattc tgaggagaat gtctgtgagg tggaactaga cacagatctc
     2881 tttccccggc cacggtcacc cagccccgaa gatgaatcca gcagttccag cagctctagc
     2941 agctctgagg atgaggagga gctgaatgaa cgccgagcct ctacctggca gcggaatgcc
     3001 atgcggcgcc gacagaagac aacccgagaa gacaagccca gtgccccaat caagcccacc
     3061 aacacttaca ttggagaaga caactatgat tacccccaga tcaaagtgga tgacctctcc
     3121 tcctccccaa cctcgtcccc tgagcggagc acttccacgc tagagattca accaagccgg
     3181 gcatcaccaa cttctgacat agaatcagtt gagcgaaaaa tttataaagc ttacaagtgg
     3241 ctccgctact cttatatctc ctactcaaat aacaaagatg gagagacctc cttggtgact
     3301 ggggaggcag atgaagggag agcaggaacc agccacaaag acaacccagc cccttcttcc
     3361 agtaaggaag cctgtctaaa catagcaatg gcccagagga accaggacct gccacctgaa
     3421 ggctgcagca aggacacttt taaagaagag actcctagaa ctcccagcaa tggcccaggc
     3481 catgagcaca gcagccatgc ttgggcagag gtgccagagg gtacctctca ggacactggc
     3541 aatagcggct ctgtagagca cccttttgaa accaagaagc tcaatggaaa ggccctgagc
     3601 agtcgggctg aggagccgcc ttctcctcct gtccccaagg catctggctc cactctcaac
     3661 agcgggtctg gcaactgtcc caggacccag tctgatgaca gtgaggagag gagcctcgaa
     3721 accatctgtg ccaaccacaa caatggacgc ttacaccctc gtccccctca ccctcacaat
     3781 aacgggcaga acttggggga gctggaggtg gtggcctact cttccccagg acactcagac
     3841 actgaccgtg ataactcgtc cctgacaggg acactcctac acaaagattg ttgcgggtct
     3901 gaaatggcct gtgagacccc caatgctgga acaagagagg accccactga caccccagcc
     3961 acagatagta gcagggctgt tcatggccac agtggcctca aaaggcaacg aattgaattg
     4021 gaagatacag attcagagaa ttcctcctca gagaagaaat taaaaacagg ctcgagcggt
     4081 gggagctccg gtgtgagcgg ctggcggctg ttcaagaaga ttagctaagt ttaaacggcc
     4141 gcgactctag actgcaggca tgcaagctga tttggctgct gccaccgctg agcaaggccg
     4201 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4261 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4321 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4381 gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcttaatt
     4441 aagacttcta atataacttc gtatagcata cattatacga agttatggtt ccactgttga
     4501 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     4561 ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4621 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4681 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4741 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4801 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4861 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4921 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4981 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5041 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5101 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5161 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5221 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5281 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5341 tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc
     5401 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5461 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5521 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5581 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5641 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5701 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5761 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5821 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5881 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5941 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6001 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6061 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
     6121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt ccggaatcgt
     6241 cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg cggccatcgt
     6301 gatggctaga gtttagccct cccacacata accagagggc agcaattcac gaatcccaac
     6361 tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat gcagatcgag
     6421 aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat ttccgatcgc
     6481 gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca ccacgagttc
     6541 tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc ggccgtcgct
     6601 agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt tgattgtagc
     6661 cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca tggtggcgct
     6721 agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     6781 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     6841 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     6901 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     6961 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     7021 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     7081 ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg ttatttcagg
     7141 ccatggtgct gcgc
//
