LOCUS       sequence-426017-        5991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR20. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5991)
  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 5991)
  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_WDR20:1-569
FEATURES             Location/Qualifiers
     source          1..5991
                     /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    3054..3175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3065..3084)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3290..3323)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3333..3380
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3412..4206
                     /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(3466..3485)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4076..4095
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4364..4952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4444..4463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5078..5126
                     /note="synthetic polyadenylation signal"
     CDS             complement(5150..5548)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5545..5554
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5579..5936)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5593..5728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5745..5936
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5893..5913
                     /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 tggcgacgga
     1261 gggaggaggg aaggagatga acgagattaa gacccaattc accacccggg aaggtctgta
     1321 caagctgctg ccgcactcgg agtacagccg gcccaaccgg gtgcccttca actcgcaggg
     1381 atccaaccct gtccgcgtct ccttcgtaaa cctcaacgac cagtctggca acggcgaccg
     1441 cctctgcttc aatgtgggcc gggagctgta cttctatatc tacaaggggg tccgcaaggc
     1501 tgctgacttg agtaaaccaa tagataaaag gatatacaaa ggaacacagc ctacttgtca
     1561 tgacttcaac cacctaacag ccacagcaga aagtgtctct ctcctagtgg gcttttccgc
     1621 aggccaagtc cagcttatag acccaatcaa aaaagaaact agcaaacttt ttaatgagga
     1681 aagactaata gacaagtcac gagttacctg tgtcaaatgg gttcacggtt cggaaagcct
     1741 tttcctagta gcccactcga gtgggaacat gtacttatat aatgtggagc acacttgtgg
     1801 caccacagcc ccccactacc agcttctgaa gcagggagag agctttgccg tgcacacttg
     1861 caagagcaaa tccacgagga accctctcct taagtggacg gtgggcgagg gggccctcaa
     1921 cgagtttgct ttctccccag atggcaagtt cttagcgtgc gtgagccagg acgggtttct
     1981 gcgggtgttc aactttgact cagtggagct gcacggtacg atgaaaagct actttggggg
     2041 cttgctgtgt gtgtgctgga gcccggatgg caagtacatc gtgacaggtg gggaggacga
     2101 cttggtgaca gtctggtcct ttgtagactg ccgagtaata gccagaggcc acgggcacaa
     2161 gtcctgggtc agtgttgtag cgtttgaccc ttataccact agtgtagaag aaggtgaccc
     2221 tatggagttt agtggcagcg atgaggactt ccaagacctt cttcattttg gcagagatcg
     2281 agcaaatagt acacagtcca ggctctccaa acggaactct acagacagcc gccccgtaag
     2341 tgtcacgtat cggtttggtt ccgtgggcca ggacacacag ctctgtttat gggaccttac
     2401 agaagatatc cttttccctc accaacccct ctcaagagca aggacacaca caaatgtcat
     2461 gaatgccacg agtcctcctg ctggaagcaa tgggaacagt gttacaacac ccgggaactc
     2521 tgtgccgcct cctctgccac ggtccaacag ccttccacat tcagcagtct caaatgctgg
     2581 cagcaaaagc agtgtcatgg acggggccat tgcttctggg gtcagcaaat ttgcaacact
     2641 ttcactacat gaccggaagg agaggcacca cgagaaagat cacaagcgaa atcatagcat
     2701 gggacacatt tctagcaaga gcagtgacaa actgaatcta gttaccaaaa ccaaaacgga
     2761 ccctgctaaa actctgggaa cgcccctgtg tcctcgaatg gaagatgttc ccttgttaga
     2821 gccgctgata tgtaaaaaga tagcacatga gagactgact gtactaatat ttcttgaaga
     2881 ctgtatagtc actgcttgtc aggagggatt tatttgcaca tggggaaggc ctggtaaagt
     2941 ggtaagtttt aatccttaat ctagaggtca agcttcgggg gtttaaacct gcaggcatgc
     3001 aagctgattt ggctgctgcc accgctgagc aaggccgctt cgagcagaca tgataagata
     3061 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3121 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3181 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg tttttttaag
     3241 caagtaaaac ctctacaaat gtggtaaaat cttaattaag acttctaata taacttcgta
     3301 tagcatacat tatacgaagt tatggttcca ctgttgacaa ttaatcatcg gcatagtata
     3361 tcggcatagt ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa
     3421 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3481 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3541 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3601 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3661 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3721 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3781 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3841 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3901 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3961 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4021 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4081 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4141 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4201 ttctgattcg aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg
     4261 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4321 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4381 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4441 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4501 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4561 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4621 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4681 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4741 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4801 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4861 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4921 agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt
     4981 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5041 ggatcttcac ctagatcctt ttatagtccg gaatcgtcac acaaaaaacc aacacacaga
     5101 tgtaatgaaa ataaagatat tttattgcgg ccatcgtgat ggctagagtt tagccctccc
     5161 acacataacc agagggcagc aattcacgaa tcccaactgc cgtcggctgt ccatcactgt
     5221 ccttcactat ggctttgatc ccaggatgca gatcgagaag cacctgtcgg caccgtccgc
     5281 aggggctcaa gatgcccctg ttctcatttc cgatcgcgac gatacaagtc aggttgccag
     5341 ctgccgcagc agcagcagtg cccagcacca cgagttctgc acaaggtccc ccagtaaaat
     5401 gatatacatt gacaccagtg aagatgcggc cgtcgctaga gagagctgcg ctggcgacgc
     5461 tgtagtcttc agagatgggg atgctgttga ttgtagccgt tgctctttca atgagggtgg
     5521 attcttcttg agacaaaggc ttggccatgg tggcgctagt gtcagaagaa tcgagctttt
     5581 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     5641 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     5701 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     5761 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     5821 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     5881 gggactttcc acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg
     5941 tacctaacca agttcctctt tcagaggtta tttcaggcca tggtgctgcg c
//
