LOCUS       sequence-423357-        5330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR5. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5330)
  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 5330)
  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-C_WDR5:1-334
FEATURES             Location/Qualifiers
     source          1..5330
                     /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    2393..2514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2404..2423)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2629..2662)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2672..2719
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2751..3545
                     /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(2805..2824)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3415..3434
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3703..4291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3783..3802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4417..4465
                     /note="synthetic polyadenylation signal"
     CDS             complement(4489..4887)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4884..4893
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4918..5275)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4932..5067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5084..5275
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5232..5252
                     /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 ataggggcga tcgcaacaag cttcgggaag
     1201 atctggagct agcggcgggg gatccggtgg gaattcgaca ccatggcgac ggaggagaag
     1261 aagcccgaga ccgaggccgc cagagcacag ccaacccctt cgtcatccgc cactcagagc
     1321 aagcctacac ctgtgaagcc aaactatgct ctaaagttca cccttgctgg ccacaccaaa
     1381 gcagtgtcct ccgtgaaatt cagcccgaat ggagagtggc tggcaagttc atctgctgat
     1441 aaacttatta aaatttgggg cgcgtatgat gggaaatttg agaaaaccat atctggtcac
     1501 aagctgggaa tatccgatgt agcctggtcg tcagattcta accttcttgt ttctgcctca
     1561 gatgacaaaa ccttgaagat atgggacgtg agctcgggca agtgtctgaa aaccctgaag
     1621 ggacacagta attatgtctt ttgctgcaac ttcaatcccc agtccaacct tattgtctca
     1681 ggatcctttg acgaaagcgt gaggatatgg gatgtgaaaa cagggaagtg cctcaagact
     1741 ttgccagctc actcggatcc agtctcggcc gttcatttta atcgtgatgg atccttgata
     1801 gtttcaagta gctatgatgg tctctgtcgc atctgggaca ccgcctcggg ccagtgcctg
     1861 aagacgctca tcgatgacga caaccccccc gtgtcttttg tgaagttctc cccgaacggc
     1921 aaatacatcc tggccgccac gctggacaac actctgaagc tctgggacta cagcaagggg
     1981 aagtgcctga agacgtacac tggccacaag aatgagaaat actgcatatt tgccaatttc
     2041 tctgttactg gtgggaagtg gattgtgtct ggctcagagg ataaccttgt ttacatctgg
     2101 aaccttcaga cgaaagagat tgtacagaaa ctacaaggcc acacagatgt cgtgatctca
     2161 acagcttgtc acccaacaga aaacatcatc gcctctgctg cgctagaaaa tgacaaaaca
     2221 attaaactgt ggaagagtga ctgcggctcg agcggtggga gctccggtgt gagcggctgg
     2281 cggctgttca agaagattag ctaagtttaa acggccgcga ctctagactg caggcatgca
     2341 agctgatttg gctgctgcca ccgctgagca aggccgcttc gagcagacat gataagatac
     2401 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2461 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2521 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc
     2581 aagtaaaacc tctacaaatg tggtaaaatc ttaattaaga cttctaatat aacttcgtat
     2641 agcatacatt atacgaagtt atggttccac tgttgacaat taatcatcgg catagtatat
     2701 cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac
     2761 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2821 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2881 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     2941 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3001 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3061 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3121 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3181 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3241 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3301 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3361 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3421 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3481 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3541 tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     3601 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4141 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4261 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     4321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4381 gatcttcacc tagatccttt tatagtccgg aatcgtcaca caaaaaacca acacacagat
     4441 gtaatgaaaa taaagatatt ttattgcggc catcgtgatg gctagagttt agccctccca
     4501 cacataacca gagggcagca attcacgaat cccaactgcc gtcggctgtc catcactgtc
     4561 cttcactatg gctttgatcc caggatgcag atcgagaagc acctgtcggc accgtccgca
     4621 ggggctcaag atgcccctgt tctcatttcc gatcgcgacg atacaagtca ggttgccagc
     4681 tgccgcagca gcagcagtgc ccagcaccac gagttctgca caaggtcccc cagtaaaatg
     4741 atatacattg acaccagtga agatgcggcc gtcgctagag agagctgcgc tggcgacgct
     4801 gtagtcttca gagatgggga tgctgttgat tgtagccgtt gctctttcaa tgagggtgga
     4861 ttcttcttga gacaaaggct tggccatggt ggcgctagtg tcagaagaat cgagcttttt
     4921 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     4981 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     5041 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     5101 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     5161 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     5221 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcagaaggt
     5281 acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc
//
