LOCUS       sequence-423402-        5544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDF3. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5544)
  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 5544)
  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_WDF3:1-420
FEATURES             Location/Qualifiers
     source          1..5544
                     /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    2607..2728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2618..2637)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2843..2876)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2886..2933
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2965..3759
                     /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(3019..3038)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3629..3648
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3917..4505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3997..4016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4631..4679
                     /note="synthetic polyadenylation signal"
     CDS             complement(4703..5101)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5098..5107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5132..5489)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5146..5281
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5298..5489
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5446..5466
                     /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 tgggcacctc
     1261 caaagaaaag gccaagcccc tcacgctcaa gcaggctctc cttggccaca ctgatactgt
     1321 cacctgtgcc agagcatccc tggcctacca cataattgtc agcgggtccc gggaccgaac
     1381 ctgcatcatc tgggacctga acaaactgtc attcctaacc cagctccgcg gccaccgagc
     1441 tcccgtctcc gctctttgca ttaatgagct aacaggggac attgtgtcct gcgcaggcac
     1501 gtacatccac gtgtggagca tcaatgggaa tcccatcgtg agtgtgaaca cattcacggg
     1561 ccggagccag cagattgtct gctgctgcat gtccgagatg aacgagtggg acacgcagaa
     1621 tgtcatcgtg acaggacact cagatggagt ggtccggttc tggagaatgg aattcctgca
     1681 ggttcctgaa acaccagctc ctgagcctgt ggaagacctg gagatgcagg agggctgccc
     1741 agaagcacag atagggcagc aagcccaaga tgacgacagc agtgattctg aaacagaaga
     1801 gcccagtgtc agccaggacc ccaaggacac gtccagccag cccagcagca ccagccacag
     1861 gccgcgggca gcctcctgca gggccaccgc cacatggtgt accgacagcg gctccgacga
     1921 ctccaggcgc tggtccgacc agctcagtct agatgagaaa gatggcttca tatttgtgaa
     1981 ctactctgag ggccagacca gagcacatct gcagggcccc ctcgcccacc cgcaccccaa
     2041 ccccatcgaa gcgcgcagtt acagcagatt gaaacccggg taccgttggg agcggcagct
     2101 ggtattcaga agcaaactga ctatgcacac agcctttgat cgaaaggaca acacgcaccc
     2161 agccgaggtc accgcgttgg gtgtatccaa ggatcacagc aggatcctcg tgggtgacag
     2221 tcgaggccga gtgttcagct ggtctgtgag tgaccagcct ggtcgttctg ccgctgacca
     2281 ctgggtgaag gatgaaggcg gcgacagctg ctcgggatgc tccgtacgct tctccctcac
     2341 agagaggagg caccattgca ggaactgtgg gcagctcttc tgtcagaagt gcagtcggtt
     2401 tcagtcagaa atcaagcgcc tgaaaatctc gtccccagtg ggtgtctgtc agaactgtta
     2461 ctatagcttg cagcatgaga ggggggcaga agatggacct cggaactgct aatctagagg
     2521 tcaagcttcg ggggtttaaa cctgcaggca tgcaagctga tttggctgct gccaccgctg
     2581 agcaaggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     2641 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2701 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2761 ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa
     2821 aatcttaatt aagacttcta atataacttc gtatagcata cattatacga agttatggtt
     2881 ccactgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     2941 ggaactaaac ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg
     3001 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3061 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3121 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3181 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3241 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3301 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3361 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3421 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3481 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3541 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3601 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3661 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3721 cgcatcgcct tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg
     3781 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3841 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3901 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3961 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4021 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4081 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4141 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4201 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4261 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4321 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4381 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4441 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4501 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4561 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     4621 ccggaatcgt cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg
     4681 cggccatcgt gatggctaga gtttagccct cccacacata accagagggc agcaattcac
     4741 gaatcccaac tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat
     4801 gcagatcgag aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat
     4861 ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca
     4921 ccacgagttc tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc
     4981 ggccgtcgct agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt
     5041 tgattgtagc cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca
     5101 tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc
     5161 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     5221 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     5281 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     5341 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     5401 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     5461 cattccacag ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg
     5521 ttatttcagg ccatggtgct gcgc
//
