LOCUS       sequence-426000-        5223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR82. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5223)
  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 5223)
  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_WDR82:1-313
FEATURES             Location/Qualifiers
     source          1..5223
                     /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    2286..2407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2297..2316)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2522..2555)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2565..2612
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2644..3438
                     /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(2698..2717)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3308..3327
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3596..4184)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3676..3695)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4310..4358
                     /note="synthetic polyadenylation signal"
     CDS             complement(4382..4780)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4777..4786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4811..5168)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4825..4960
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4977..5168
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5125..5145
                     /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 tgaagctgac
     1261 cgacagcgtg ttgcggagct tccgcgtcgc taaggtgttc cgcgaaaact cggacaagat
     1321 taactgcttc gatttcagcc ccaacggcga gacggtcatc tcgagtagcg acgacgactc
     1381 catcgtgctc tatgactgcc aggagggcaa accaaagaga accctgtaca gtaagaaata
     1441 tggtgtggac ctcatcagat acactcatgc agcaaacaca gttgtttaca gctctaacaa
     1501 aatagacgat actattcgtt acttgtcctt gcatgacaac aaatacatca gatactttcc
     1561 tggacatagc aaaagggtgg tggccttgtc catgtcacct gtggatgaca ctttcatttc
     1621 tgggtctctt gataagacca ttcgactctg ggatctccgg tctcctaact gccagggcct
     1681 catgcatctg caggggaagc cagtttgttc ttttgatcca gaagggttaa ttttcgctgc
     1741 aggtgtcaac tctgaaatgg tcaagcttta tgaccttcgt tcttttgata aggggccatt
     1801 tgctaccttt aagatgcagt atgatcgaac ttgtgagtgg acaggactta aattcagcaa
     1861 tgatggcaag ctcatcctca tttccaccaa cggcagcttc attcgtctga ttgatgcatt
     1921 caaaggagtg gtgatgcaca catttggggg ttatgccaac agcaaagctg tcacactgga
     1981 ggcttcattt actccagact ctcagtttat tatgattggt tcagaggatg gcaagatcca
     2041 tgtctggaat ggagagagcg gtataaaagt agctgtgttg gatggtaaac acacaggccc
     2101 gattacctgt ttgcaattca accccaagtt catgactttt gccagtgcgt gttccaacat
     2161 ggccttttgg ttgcccacca ttgatgacta atctagaggt caagcttcgg gggtttaaac
     2221 ctgcaggcat gcaagctgat ttggctgctg ccaccgctga gcaaggccgc ttcgagcaga
     2281 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2341 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2401 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     2461 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcttaatta agacttctaa
     2521 tataacttcg tatagcatac attatacgaa gttatggttc cactgttgac aattaatcat
     2581 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag
     2641 atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2701 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2761 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2821 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2881 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2941 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3001 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3061 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3121 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3181 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3241 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3301 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3361 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3421 cttgacgagt tcttctgatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc
     3481 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3541 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3601 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3661 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3721 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3781 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3841 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3901 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3961 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4021 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4081 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4141 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     4201 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4261 gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaatcgtc acacaaaaaa
     4321 ccaacacaca gatgtaatga aaataaagat attttattgc ggccatcgtg atggctagag
     4381 tttagccctc ccacacataa ccagagggca gcaattcacg aatcccaact gccgtcggct
     4441 gtccatcact gtccttcact atggctttga tcccaggatg cagatcgaga agcacctgtc
     4501 ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt tccgatcgcg acgatacaag
     4561 tcaggttgcc agctgccgca gcagcagcag tgcccagcac cacgagttct gcacaaggtc
     4621 ccccagtaaa atgatataca ttgacaccag tgaagatgcg gccgtcgcta gagagagctg
     4681 cgctggcgac gctgtagtct tcagagatgg ggatgctgtt gattgtagcc gttgctcttt
     4741 caatgagggt ggattcttct tgagacaaag gcttggccat ggtggcgcta gtgtcagaag
     4801 aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     4861 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     4921 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     4981 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     5041 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     5101 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc tggttctttc
     5161 cgcctcagaa ggtacctaac caagttcctc tttcagaggt tatttcaggc catggtgctg
     5221 cgc
//
