LOCUS       sequence-423410-        5412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR86. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5412)
  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 5412)
  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_WDR86:1-376
FEATURES             Location/Qualifiers
     source          1..5412
                     /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    2475..2596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2486..2505)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2711..2744)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2754..2801
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2833..3627
                     /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(2887..2906)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3497..3516
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3785..4373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3865..3884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4499..4547
                     /note="synthetic polyadenylation signal"
     CDS             complement(4571..4969)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4966..4975
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5000..5357)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5014..5149
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5166..5357
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5314..5334
                     /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 tgggaggtgg
     1261 aggatctgca ctgagggtat gtgcggacca tcgaggggga atcaattggt tgtccttgtc
     1321 cccagacggc caacgcctgc tgacaggctc agaagatgga acggctcggc tctggtccac
     1381 agcagacggt cagtgttgtg cactgttgca gggacacgag tcatatgtca cattttgcca
     1441 gttggaggat gaggccgctt tcacttgtag cgcggattgt accataaggc ggtgggatgt
     1501 gttgaccggt cagtgcttgc aggtttacag aggacatacc agcatagtca atcggatcct
     1561 cgtcgccaac aaccagctct tcagtagttc ctatgaccga acagcacggg tctggagcgt
     1621 tgataaggga caaatgtccc gagaattcag ggggcaccgc aactgtgttc ttacactggc
     1681 ctactcagca ccgtgggatt tgccatcaac cccctgtgct gaagaagcgg cagctggggg
     1741 gctcctggtc acggggtcaa ctgacggtac cgcaaaagtt tggcaggtcg catcagggtg
     1801 ttgtcaccag acactgcgcg gtcacacggg ggcggttctt tgcttggtgc tcgatacccc
     1861 aggacacacg gcatttacag gatccactga cgctacgata cgcgcgtggg acatcttgag
     1921 tggtgagcaa ctgagagtgt ttagagagca tcgaggttca gtgatttgtc tggaattggt
     1981 aaaccggttg gtttacagcg gttccgctga caggacagtg aagtgttggt tggcggatac
     2041 aggagagtgt gtaaggacat tcactgccca cagacggaac gtatcagctc ttaaatatca
     2101 cgctgggact ctgtttactg ggtctgggga cgcgtgcgcc agggcttttg atgcacaaag
     2161 tggtgagctg agaagggtat ttagggggca cacctttatc attaactgta tacaagtgca
     2221 tggtcaagtg ctttacactg ctagccatga tggtgccctc cgactgtggg acgttcgagg
     2281 tcttaggggt gctcccagac ctccaccccc tatgagatct ctgagcagac tgttctcaaa
     2341 taaggtaggg tgtgctgccg cacccctcca accagcctaa tctagaggtc aagcttcggg
     2401 ggtttaaacc tgcaggcatg caagctgatt tggctgctgc caccgctgag caaggccgct
     2461 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2521 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2581 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2641 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcttaattaa
     2701 gacttctaat ataacttcgt atagcataca ttatacgaag ttatggttcc actgttgaca
     2761 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc
     2821 aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     2881 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     2941 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3001 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3061 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3121 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3181 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3241 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3301 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3361 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3421 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3481 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3541 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3601 tatcgccttc ttgacgagtt cttctgattc gaaatgaccg accaagcgac gcccaaccgg
     3661 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3721 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3781 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3841 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3901 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3961 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4021 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4081 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4141 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4201 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4261 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4321 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     4381 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4441 tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc ggaatcgtca
     4501 cacaaaaaac caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga
     4561 tggctagagt ttagccctcc cacacataac cagagggcag caattcacga atcccaactg
     4621 ccgtcggctg tccatcactg tccttcacta tggctttgat cccaggatgc agatcgagaa
     4681 gcacctgtcg gcaccgtccg caggggctca agatgcccct gttctcattt ccgatcgcga
     4741 cgatacaagt caggttgcca gctgccgcag cagcagcagt gcccagcacc acgagttctg
     4801 cacaaggtcc cccagtaaaa tgatatacat tgacaccagt gaagatgcgg ccgtcgctag
     4861 agagagctgc gctggcgacg ctgtagtctt cagagatggg gatgctgttg attgtagccg
     4921 ttgctctttc aatgagggtg gattcttctt gagacaaagg cttggccatg gtggcgctag
     4981 tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     5041 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     5101 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     5161 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5221 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     5281 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct
     5341 ggttctttcc gcctcagaag gtacctaacc aagttcctct ttcagaggtt atttcaggcc
     5401 atggtgctgc gc
//
