LOCUS       sequence-426011-        5310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR77. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5310)
  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 5310)
  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_WDR77:1-342
FEATURES             Location/Qualifiers
     source          1..5310
                     /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    2373..2494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2384..2403)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2609..2642)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2652..2699
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2731..3525
                     /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(2785..2804)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3395..3414
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3683..4271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3763..3782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4397..4445
                     /note="synthetic polyadenylation signal"
     CDS             complement(4469..4867)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4864..4873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4898..5255)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4912..5047
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5064..5255
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5212..5232
                     /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 tgcggaagga
     1261 aaccccaccc cccctagtgc ccccggcggc ccgggagtgg aatcttcccc caaatgcgcc
     1321 cgcctgcatg gaacggcagt tggaggctgc gcggtaccgg tccgatgggg cgcttctcct
     1381 cggggcctcc agcctgagtg ggcgctgctg ggccggctcc ctctggcttt ttaaggaccc
     1441 ctgtgccgcc cccaacgaag gcttctgctc cgccggagtc caaacggagg ctggagtggc
     1501 tgacctcact tgggttgggg agagaggtat tctagtggcc tccgattcag gtgctgttga
     1561 attgtgggaa ctagatgaga atgagacact tattgtcagc aagttctgca agtatgagca
     1621 tgatgacatt gtgtctacag tcagtgtctt gagctctggc acacaagctg tcagtggtag
     1681 caaagacatc tgcatcaagg tttgggacct tgctcagcag gtggtactga gttcataccg
     1741 agctcatgct gctcaggtca cttgtgttgc tgcctctcct cacaaggact ctgtgtttct
     1801 ttcatgcagc gaggacaata gaattttact ctgggatacc cgctgtccca agccagcatc
     1861 acagattggc tgcagtgcgc ctggctacct tcctacctcg ctggcttggc atcctcagca
     1921 aagtgaagtc tttgtctttg gtgatgagaa tgggacagtc tcccttgtgg acaccaagag
     1981 tacaagctgt gtcctgagct cagctgtaca ctcccagtgt gtcactgggc tggtgttctc
     2041 cccacacagt gttcccttcc tggcctctct cagtgaagac tgctcacttg ctgtgctgga
     2101 ctcaagcctt tctgagttgt ttagaagcca agcccacaga gactttgtga gagatgcgac
     2161 ttggtccccg ctcaatcact ccctgcttac cacagtgggc tgggaccatc aggtcgtcca
     2221 ccacgttgtg cccacagaac ctctcccagc ccctggacct gcaagtgtta ctgagtaatc
     2281 tagaggtcaa gcttcggggg tttaaacctg caggcatgca agctgatttg gctgctgcca
     2341 ccgctgagca aggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2401 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2461 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2521 gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg
     2581 tggtaaaatc ttaattaaga cttctaatat aacttcgtat agcatacatt atacgaagtt
     2641 atggttccac tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     2701 aggtgaggaa ctaaacccag gaggcagatc atgattgaac aagatggatt gcacgcaggt
     2761 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2821 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     2881 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     2941 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3001 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3061 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3121 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3181 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3241 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3301 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3361 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3421 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3481 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgattcga aatgaccgac
     3541 caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3601 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3661 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3721 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3781 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3841 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3901 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3961 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4021 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4081 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4141 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4201 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4261 aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4321 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4381 tatagtccgg aatcgtcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt
     4441 ttattgcggc catcgtgatg gctagagttt agccctccca cacataacca gagggcagca
     4501 attcacgaat cccaactgcc gtcggctgtc catcactgtc cttcactatg gctttgatcc
     4561 caggatgcag atcgagaagc acctgtcggc accgtccgca ggggctcaag atgcccctgt
     4621 tctcatttcc gatcgcgacg atacaagtca ggttgccagc tgccgcagca gcagcagtgc
     4681 ccagcaccac gagttctgca caaggtcccc cagtaaaatg atatacattg acaccagtga
     4741 agatgcggcc gtcgctagag agagctgcgc tggcgacgct gtagtcttca gagatgggga
     4801 tgctgttgat tgtagccgtt gctctttcaa tgagggtgga ttcttcttga gacaaaggct
     4861 tggccatggt ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa
     4921 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     4981 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     5041 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     5101 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     5161 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     5221 gacacacatt ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt
     5281 cagaggttat ttcaggccat ggtgctgcgc
//
