LOCUS       sequence-423403-        5916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR25. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5916)
  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 5916)
  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_WDR25:1-544
FEATURES             Location/Qualifiers
     source          1..5916
                     /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    2979..3100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2990..3009)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3215..3248)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3258..3305
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3337..4131
                     /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(3391..3410)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4001..4020
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4289..4877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4369..4388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5003..5051
                     /note="synthetic polyadenylation signal"
     CDS             complement(5075..5473)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5470..5479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5504..5861)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5518..5653
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5670..5861
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5818..5838
                     /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 tgacagcaag
     1261 aactctgtct ttaatggctt cattggtagc gtatgatgat tcggactcgg aggctgagac
     1321 agagcatgca ggaagtttta atgctaccgg ccagcagaaa gacacttctg gtgtggccag
     1381 accacctggg caggattttg catctggtac actggatgtg cccaaagcag gggcacagcc
     1441 cacaaagcat ggctcctgtg aagacccagg gggctatcgc cttccattgg ctcagcttgg
     1501 gagaagcgat tggggatctt gccccagcca gaggctacag tggcccggga aggagcctca
     1561 agtcaccttc cccatcaaag agccttcttg ttcttctctg tggacgagcc atgttccagc
     1621 cagccacatg cccctggcag ctgcccgctt taagcaagta aaactctcca ggaactttcc
     1681 caagtcatct ttccatgctc aaagtgagtc tgaaaccgta ggtaaaaatg gcagctcttt
     1741 tcagaagaaa aaatgtgagg actgtgtggt accctatact cccagaagac taagacagcg
     1801 gcaggcatta agcacggaga caggcaaggg taaagacgtg gagccacagg ggccccctgc
     1861 agggcgtgcc ccagcccctc tctacgtggg cccgggagtg tctgagttta ttcagccata
     1921 tttgaatagc cattataaag aaaccacagt tccccggaaa gtgcttttcc acctgagagg
     1981 ccacaggggc cctgtcaaca ccattcagtg gtgtccagtc ctttctaaga gccacatgct
     2041 tctctccact tctatggata aaactttcaa ggtatggaac gccgtggact ccgggcactg
     2101 cctgcagacc tactccctgc acacagaggc agtgcgggcc gcccggtggg ctccctgtgg
     2161 ccggcgcatc ctcagtggtg gctttgactt cgcgctgcac ctaacagacc ttgaaacagg
     2221 aacccagcta tttagtggtc gaagtgactt tagaatcact accttgaaat tccatccaaa
     2281 agaccacaac atctttttat gtggaggctt cagctctgaa atgaaagctt gggatataag
     2341 gactggcaag gtgatgagaa gctacaaggc gaccatccag cagaccttgg acatcctgtt
     2401 cctccgggaa ggctccgagt tcctgagcag cacagacgct tccacccggg actcagctga
     2461 ccgcaccatt attgcctggg atttccggac ctctgccaaa atctccaacc agattttcca
     2521 cgagaggttc acctgcccca gcctcgcctt gcacccgaga gagcccgtgt tcctggcaca
     2581 gaccaatggc aactacctgg cccttttctc cactgtgtgg ccctaccgga tgagcagacg
     2641 gcggcgctat gaagggcaca aggtggaggg ctactcagtg ggctgcgagt gctccccagg
     2701 cggtgacttg ctggtgacgg gcagcgccga tggccgggtc ctgatgtaca gcttccgcac
     2761 agccagccga gcatgcacac tgcaggggca cacacaggcc tgtgtcggca ccacctatca
     2821 ccccgtgctg ccctccgtcc tcgccacctg ctcctgggga ggggacatga agatctggca
     2881 ctaatctaga ggtcaagctt cgggggttta aacctgcagg catgcaagct gatttggctg
     2941 ctgccaccgc tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3001 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3061 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3121 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     3181 caaatgtggt aaaatcttaa ttaagacttc taatataact tcgtatagca tacattatac
     3241 gaagttatgg ttccactgtt gacaattaat catcggcata gtatatcggc atagtataat
     3301 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     3361 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3421 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3481 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3541 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3601 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3661 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3721 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3781 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3841 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3901 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3961 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4021 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4081 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg
     4141 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     4201 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4261 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4861 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4921 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4981 tccttttata gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa
     5041 gatattttat tgcggccatc gtgatggcta gagtttagcc ctcccacaca taaccagagg
     5101 gcagcaattc acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt
     5161 tgatcccagg atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc
     5221 ccctgttctc atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag
     5281 cagtgcccag caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac
     5341 cagtgaagat gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga
     5401 tggggatgct gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca
     5461 aaggcttggc catggtggcg ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc
     5521 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     5581 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     5641 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     5701 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
     5761 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     5821 ctaactgaca cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc
     5881 ctctttcaga ggttatttca ggccatggtg ctgcgc
//
