LOCUS       sequence-423407-        5358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR53. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5358)
  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 5358)
  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_WDR53:1-358
FEATURES             Location/Qualifiers
     source          1..5358
                     /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    2421..2542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2432..2451)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2657..2690)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2700..2747
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2779..3573
                     /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(2833..2852)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3443..3462
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3731..4319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3811..3830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4445..4493
                     /note="synthetic polyadenylation signal"
     CDS             complement(4517..4915)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4912..4921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4946..5303)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4960..5095
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5112..5303
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5260..5280
                     /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 tggcagtcaa
     1261 gtggacgggt gggcattctt ctcctgtcct ctgcctgaat gcaagtaaag aagggctgct
     1321 ggcttctgga gcagagggcg gagatctcac ggcttggggt gaagatggaa ctccattagg
     1381 acacacgcgg ttccaagggg ctgatgatgt taccagtgtc ttattttctc cctcctgtcc
     1441 caccaagctc tatgcctcac atggagaaac cattagtgta ctggatgtca ggtccctcaa
     1501 agattccttg gaccattttc atgtgaatga agaagaaatc aattgtcttt cattgaatca
     1561 aacggaaaac ctgctggctt ctgctgacga ctctggggca atcaaaatcc tagacttgga
     1621 aaacaagaaa gttatcagat ccttgaagag acattccaat atctgctcct cagtggcttt
     1681 tcggcctcag aggcctcaga gcctggtgtc atgtggactg gatatgcagg tgatgctgtg
     1741 gagtcttcaa aaagcccgac cactctggat tacaaattta caggaggatg aaacagaaga
     1801 aatggaaggc ccacagtcac ctggtcagct cttaaaccct gccctagccc attctatctc
     1861 tgtggcttcg tgtggtaata tttttagttg tggtgcagaa gatggtaagg ttcgaatctt
     1921 tcgggtgatg ggagttaagt gtgaacagga actgggattt aagggccaca cttcaggggt
     1981 atcccaggtc tgctttctcc cagaatccta tttgctgctt actggaggga atgatgggaa
     2041 gatcacgttg tgggatgcaa acagtgaagt tgagaaaaaa cagaagagtc ccacaaaacg
     2101 tacccacagg aagaaaccta aaagaggaac ttgcaccaag cagggtggaa atactaacgc
     2161 ttcagtaaca gatgaggaag aacatggcaa cattttacca aagctaaata ttgaacatgg
     2221 agaaaaagtg aactggctct tgggtacaaa aataaaggga caccaaaata tattagtagc
     2281 tgatcaaact agttgtatat ctgtataccc cttaaatgaa ttttaatcta gaggtcaagc
     2341 ttcgggggtt taaacctgca ggcatgcaag ctgatttggc tgctgccacc gctgagcaag
     2401 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2461 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2521 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2581 gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatctt
     2641 aattaagact tctaatataa cttcgtatag catacattat acgaagttat ggttccactg
     2701 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     2761 aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2821 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2881 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2941 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3001 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3061 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3121 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3181 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3241 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3301 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3361 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3421 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3481 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3541 gccttctatc gccttcttga cgagttcttc tgattcgaaa tgaccgacca agcgacgccc
     3601 aaccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3661 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3721 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3781 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3841 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3901 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3961 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4021 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4081 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4141 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4201 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4261 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag
     4321 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4381 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa
     4441 tcgtcacaca aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca
     4501 tcgtgatggc tagagtttag ccctcccaca cataaccaga gggcagcaat tcacgaatcc
     4561 caactgccgt cggctgtcca tcactgtcct tcactatggc tttgatccca ggatgcagat
     4621 cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat gcccctgttc tcatttccga
     4681 tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc agcagtgccc agcaccacga
     4741 gttctgcaca aggtccccca gtaaaatgat atacattgac accagtgaag atgcggccgt
     4801 cgctagagag agctgcgctg gcgacgctgt agtcttcaga gatggggatg ctgttgattg
     4861 tagccgttgc tctttcaatg agggtggatt cttcttgaga caaaggcttg gccatggtgg
     4921 cgctagtgtc agaagaatcg agctttttgc aaaagcctag gcctccaaaa aagcctcctc
     4981 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     5041 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     5101 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     5161 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
     5221 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     5281 acagctggtt ctttccgcct cagaaggtac ctaaccaagt tcctctttca gaggttattt
     5341 caggccatgg tgctgcgc
//
