LOCUS       sequence-423409-        5253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human WDR81 fragment A1619 to A1941. 
            N-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5253)
  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 5253)
  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_WDR81:1619-1941
FEATURES             Location/Qualifiers
     source          1..5253
                     /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    2316..2437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2327..2346)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2552..2585)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2595..2642
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2674..3468
                     /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(2728..2747)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3338..3357
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3626..4214)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3706..3725)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4340..4388
                     /note="synthetic polyadenylation signal"
     CDS             complement(4412..4810)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4807..4816
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4841..5198)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4855..4990
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5007..5198
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5155..5175
                     /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 tcgagcggtg cgtactggca
     1261 gtacgagatc ggcgtgagcc agcaggatgc ccactttcac ttccaccaga tccgcctgca
     1321 gagcttcccg ggccactcgg gggccgtcaa gtgcgtggca cccctgagca gcgaggactt
     1381 cttcctgagc ggcagcaagg atcgtaccgt gcgcctctgg ccgctgtaca actacggcga
     1441 cgggaccagc gagacggccc cacgcctcgt ctacacccag caccgcaaga gcgtcttctt
     1501 cgtgggccag cttgaggccc cacagcacgt ggtgagctgt gacggggctg tgcacgtctg
     1561 ggaccccttc acagggaaga cccttcgcac agtggagccg ctggacagcc gggtgcccct
     1621 gactgcggtg gctgtcatgc ccgcccccca caccagcatc accatggcca gctctgactc
     1681 taccctgcgc tttgtggact gcaggaagcc tggtctgcag cacgagttcc gactgggcgg
     1741 tgggctgaac cctgggcttg tccgtgccct ggccatcagc cccagtggcc gtagtgtcgt
     1801 ggccggcttc tcctcaggct tcatggtgct cctggacacc cgcacgggcc tggttctgcg
     1861 aggctggcca gcccacgagg gggacattct gcagatcaag gcggtggagg gcagcgtcct
     1921 ggtcagctcc tcctctgacc attccttgac cgtctggaag gagctggagc agaagcccac
     1981 ccatcactac aagtcagcat ccgaccccat ccacaccttt gacctgtacg gcagtgaggt
     2041 ggtcactggc accgtgtcca acaagattgg cgtctgctcc ctgcttgagc caccctcgca
     2101 ggccaccacg aagctcagct ctgagaactt ccgcggcacg ctcaccagcc tggccttgct
     2161 gcccactaaa cgccacctcc tgctgggctc agacaacggg gttatccgcc tcctggcata
     2221 atctagaggt caagcttcgg gggtttaaac ctgcaggcat gcaagctgat ttggctgctg
     2281 ccaccgctga gcaaggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     2341 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2401 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2461 tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa
     2521 atgtggtaaa atcttaatta agacttctaa tataacttcg tatagcatac attatacgaa
     2581 gttatggttc cactgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     2641 acaaggtgag gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca
     2701 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     2761 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     2821 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     2881 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     2941 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3001 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3061 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3121 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3181 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     3241 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3301 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3361 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3421 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaaatgacc
     3481 gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3541 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3601 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3661 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3721 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3781 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3841 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3901 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3961 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4021 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4081 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4141 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4201 aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4261 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4321 ttttatagtc cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat
     4381 attttattgc ggccatcgtg atggctagag tttagccctc ccacacataa ccagagggca
     4441 gcaattcacg aatcccaact gccgtcggct gtccatcact gtccttcact atggctttga
     4501 tcccaggatg cagatcgaga agcacctgtc ggcaccgtcc gcaggggctc aagatgcccc
     4561 tgttctcatt tccgatcgcg acgatacaag tcaggttgcc agctgccgca gcagcagcag
     4621 tgcccagcac cacgagttct gcacaaggtc ccccagtaaa atgatataca ttgacaccag
     4681 tgaagatgcg gccgtcgcta gagagagctg cgctggcgac gctgtagtct tcagagatgg
     4741 ggatgctgtt gattgtagcc gttgctcttt caatgagggt ggattcttct tgagacaaag
     4801 gcttggccat ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc
     4861 caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg
     4921 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     4981 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
     5041 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     5101 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     5161 actgacacac attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc
     5221 tttcagaggt tatttcaggc catggtgctg cgc
//
