LOCUS       sequence-423411-        5445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR89. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5445)
  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 5445)
  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_WDR89:1-387
FEATURES             Location/Qualifiers
     source          1..5445
                     /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    2508..2629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2519..2538)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2744..2777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2787..2834
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2866..3660
                     /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(2920..2939)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3530..3549
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3818..4406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3898..3917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4532..4580
                     /note="synthetic polyadenylation signal"
     CDS             complement(4604..5002)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4999..5008
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5033..5390)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5047..5182
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5199..5390
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5347..5367
                     /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 tggaaaagat
     1261 tgaggaacaa tttgctaatc tgcacattgt taaatgttcc ttaggaacca aagagcccac
     1321 ttaccttctt ggtatagaca catcaaagac tgtccaagca ggaaaggaaa acttggttgc
     1381 tgttttatgt tctaatggat caatcagaat atatgataaa gaaaggttaa atgtactacg
     1441 agaatttagt ggatatcctg gacttcttaa tggagtcaga tttgcaaatt cctgtgacag
     1501 tgtatattca gcatgtactg atggcactgt gaaatgctgg gatgctcgag tagccagaga
     1561 aaaacctgtt cagctcttca agggttaccc ttccaatatt tttatcagtt ttgatattaa
     1621 ttgtaatgat catattattt gtgctggtac agaaaaagtt gatgatgatg cattgttggt
     1681 gttttgggat gcaaggatga attctcagaa tttatctaca actaaagact cacttggtgc
     1741 atattcagag acacatagtg atgatgtcac tcaagtacgt ttccatccca gcaatcccaa
     1801 catggtagtc tcaggttcat ctgatggcct ggtaaatgta tttgatatta atattgataa
     1861 tgaggaggat gcactggtta caacctgtaa ctcaatttca tcagtaagct gtattggttg
     1921 gtctgggaaa ggttataaac agatttactg catgacacat gatgaaggat tttattggtg
     1981 ggatcttaat catctggaca ctgatgaacc agttacacgt ttgaacatcc aggatgtcag
     2041 agaagtagtt aacatgaaag aagatgcttt ggactatttg attggtggcc tatatcatga
     2101 aaagacagac acattgcatg ttattggagg aacaaacaaa ggaaggattc atttgatgaa
     2161 ctgcagcatg tcaggactga cccatgtgac tagccttcag ggagggcatg ctgctacagt
     2221 ccgttctttc tgttggaatg tgcaagatga ttctttgttg actggaggag aagatgcaca
     2281 gttgttactt tggaaacctg gagctataga gaagaccttt acaaagaaag agagtatgaa
     2341 aatagcatcc tctgtgcacc aacgagtacg agttcatagt aatgattctt ataaaagaag
     2401 gaaaaagcag taatctagag gtcaagcttc gggggtttaa acctgcaggc atgcaagctg
     2461 atttggctgc tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga
     2521 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2581 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2641 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     2701 aaacctctac aaatgtggta aaatcttaat taagacttct aatataactt cgtatagcat
     2761 acattatacg aagttatggt tccactgttg acaattaatc atcggcatag tatatcggca
     2821 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat
     2881 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2941 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3001 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3061 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3121 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3181 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3241 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3301 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3361 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3421 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3481 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3541 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3601 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3661 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     3721 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3781 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3841 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3901 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3961 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4021 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4081 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4141 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4201 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4261 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4321 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4381 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     4441 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4501 tcacctagat ccttttatag tccggaatcg tcacacaaaa aaccaacaca cagatgtaat
     4561 gaaaataaag atattttatt gcggccatcg tgatggctag agtttagccc tcccacacat
     4621 aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca
     4681 ctatggcttt gatcccagga tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc
     4741 tcaagatgcc cctgttctca tttccgatcg cgacgataca agtcaggttg ccagctgccg
     4801 cagcagcagc agtgcccagc accacgagtt ctgcacaagg tcccccagta aaatgatata
     4861 cattgacacc agtgaagatg cggccgtcgc tagagagagc tgcgctggcg acgctgtagt
     4921 cttcagagat ggggatgctg ttgattgtag ccgttgctct ttcaatgagg gtggattctt
     4981 cttgagacaa aggcttggcc atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa
     5041 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     5101 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     5161 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     5221 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     5281 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     5341 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag aaggtaccta
     5401 accaagttcc tctttcagag gttatttcag gccatggtgc tgcgc
//
