LOCUS       sequence-423470-        5960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR25. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5960)
  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 5960)
  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_WDR25:1-544
FEATURES             Location/Qualifiers
     source          1..5960
                     /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    3023..3144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3034..3053)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3259..3292)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3302..3349
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3381..4175
                     /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(3435..3454)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4045..4064
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4333..4921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4413..4432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5047..5095
                     /note="synthetic polyadenylation signal"
     CDS             complement(5119..5517)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5514..5523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5548..5905)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5562..5697
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5714..5905
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5862..5882
                     /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 ccatgacagc aagaactctg
     1261 tctttaatgg cttcattggt agcgtatgat gattcggact cggaggctga gacagagcat
     1321 gcaggaagtt ttaatgctac cggccagcag aaagacactt ctggtgtggc cagaccacct
     1381 gggcaggatt ttgcatctgg tacactggat gtgcccaaag caggggcaca gcccacaaag
     1441 catggctcct gtgaagaccc agggggctat cgccttccat tggctcagct tgggagaagc
     1501 gattggggat cttgccccag ccagaggcta cagtggcccg ggaaggagcc tcaagtcacc
     1561 ttccccatca aagagccttc ttgttcttct ctgtggacga gccatgttcc agccagccac
     1621 atgcccctgg cagctgcccg ctttaagcaa gtaaaactct ccaggaactt tcccaagtca
     1681 tctttccatg ctcaaagtga gtctgaaacc gtaggtaaaa atggcagctc ttttcagaag
     1741 aaaaaatgtg aggactgtgt ggtaccctat actcccagaa gactaagaca gcggcaggca
     1801 ttaagcacgg agacaggcaa gggtaaagac gtggagccac aggggccccc tgcagggcgt
     1861 gccccagccc ctctctacgt gggcccggga gtgtctgagt ttattcagcc atatttgaat
     1921 agccattata aagaaaccac agttccccgg aaagtgcttt tccacctgag aggccacagg
     1981 ggccctgtca acaccattca gtggtgtcca gtcctttcta agagccacat gcttctctcc
     2041 acttctatgg ataaaacttt caaggtatgg aacgccgtgg actccgggca ctgcctgcag
     2101 acctactccc tgcacacaga ggcagtgcgg gccgcccggt gggctccctg tggccggcgc
     2161 atcctcagtg gtggctttga cttcgcgctg cacctaacag accttgaaac aggaacccag
     2221 ctatttagtg gtcgaagtga ctttagaatc actaccttga aattccatcc aaaagaccac
     2281 aacatctttt tatgtggagg cttcagctct gaaatgaaag cttgggatat aaggactggc
     2341 aaggtgatga gaagctacaa ggcgaccatc cagcagacct tggacatcct gttcctccgg
     2401 gaaggctccg agttcctgag cagcacagac gcttccaccc gggactcagc tgaccgcacc
     2461 attattgcct gggatttccg gacctctgcc aaaatctcca accagatttt ccacgagagg
     2521 ttcacctgcc ccagcctcgc cttgcacccg agagagcccg tgttcctggc acagaccaat
     2581 ggcaactacc tggccctttt ctccactgtg tggccctacc ggatgagcag acggcggcgc
     2641 tatgaagggc acaaggtgga gggctactca gtgggctgcg agtgctcccc aggcggtgac
     2701 ttgctggtga cgggcagcgc cgatggccgg gtcctgatgt acagcttccg cacagccagc
     2761 cgagcatgca cactgcaggg gcacacacag gcctgtgtcg gcaccaccta tcaccccgtg
     2821 ctgccctccg tcctcgccac ctgctcctgg ggaggggaca tgaagatctg gcacggctcg
     2881 agcggtggga gctccggtgt gagcggctgg cggctgttca agaagattag ctaagtttaa
     2941 acggccgcga ctctagactg caggcatgca agctgatttg gctgctgcca ccgctgagca
     3001 aggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3061 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3121 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3181 cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc
     3241 ttaattaaga cttctaatat aacttcgtat agcatacatt atacgaagtt atggttccac
     3301 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     3361 ctaaacccag gaggcagatc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3421 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3481 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3541 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3601 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3661 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3721 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3781 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3841 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3901 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3961 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4021 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4081 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4141 tcgccttcta tcgccttctt gacgagttct tctgattcga aatgaccgac caagcgacgc
     4201 ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4261 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4321 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4381 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4441 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4501 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4561 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4621 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4681 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4741 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4801 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4861 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca
     4921 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4981 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg
     5041 aatcgtcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt ttattgcggc
     5101 catcgtgatg gctagagttt agccctccca cacataacca gagggcagca attcacgaat
     5161 cccaactgcc gtcggctgtc catcactgtc cttcactatg gctttgatcc caggatgcag
     5221 atcgagaagc acctgtcggc accgtccgca ggggctcaag atgcccctgt tctcatttcc
     5281 gatcgcgacg atacaagtca ggttgccagc tgccgcagca gcagcagtgc ccagcaccac
     5341 gagttctgca caaggtcccc cagtaaaatg atatacattg acaccagtga agatgcggcc
     5401 gtcgctagag agagctgcgc tggcgacgct gtagtcttca gagatgggga tgctgttgat
     5461 tgtagccgtt gctctttcaa tgagggtgga ttcttcttga gacaaaggct tggccatggt
     5521 ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     5581 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     5641 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     5701 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     5761 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     5821 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     5881 ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt cagaggttat
     5941 ttcaggccat ggtgctgcgc
//
