LOCUS       sequence-425868-        5354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR77. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5354)
  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 5354)
  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_WDR77:1-342
FEATURES             Location/Qualifiers
     source          1..5354
                     /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    2417..2538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2428..2447)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2653..2686)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2696..2743
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2775..3569
                     /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(2829..2848)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3439..3458
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3727..4315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3807..3826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4441..4489
                     /note="synthetic polyadenylation signal"
     CDS             complement(4513..4911)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4908..4917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4942..5299)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4956..5091
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5108..5299
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5256..5276
                     /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 ccatgcggaa ggaaacccca
     1261 ccccccctag tgcccccggc ggcccgggag tggaatcttc ccccaaatgc gcccgcctgc
     1321 atggaacggc agttggaggc tgcgcggtac cggtccgatg gggcgcttct cctcggggcc
     1381 tccagcctga gtgggcgctg ctgggccggc tccctctggc tttttaagga cccctgtgcc
     1441 gcccccaacg aaggcttctg ctccgccgga gtccaaacgg aggctggagt ggctgacctc
     1501 acttgggttg gggagagagg tattctagtg gcctccgatt caggtgctgt tgaattgtgg
     1561 gaactagatg agaatgagac acttattgtc agcaagttct gcaagtatga gcatgatgac
     1621 attgtgtcta cagtcagtgt cttgagctct ggcacacaag ctgtcagtgg tagcaaagac
     1681 atctgcatca aggtttggga ccttgctcag caggtggtac tgagttcata ccgagctcat
     1741 gctgctcagg tcacttgtgt tgctgcctct cctcacaagg actctgtgtt tctttcatgc
     1801 agcgaggaca atagaatttt actctgggat acccgctgtc ccaagccagc atcacagatt
     1861 ggctgcagtg cgcctggcta ccttcctacc tcgctggctt ggcatcctca gcaaagtgaa
     1921 gtctttgtct ttggtgatga gaatgggaca gtctcccttg tggacaccaa gagtacaagc
     1981 tgtgtcctga gctcagctgt acactcccag tgtgtcactg ggctggtgtt ctccccacac
     2041 agtgttccct tcctggcctc tctcagtgaa gactgctcac ttgctgtgct ggactcaagc
     2101 ctttctgagt tgtttagaag ccaagcccac agagactttg tgagagatgc gacttggtcc
     2161 ccgctcaatc actccctgct taccacagtg ggctgggacc atcaggtcgt ccaccacgtt
     2221 gtgcccacag aacctctccc agcccctgga cctgcaagtg ttactgaggg ctcgagcggt
     2281 gggagctccg gtgtgagcgg ctggcggctg ttcaagaaga ttagctaagt ttaaacggcc
     2341 gcgactctag actgcaggca tgcaagctga tttggctgct gccaccgctg agcaaggccg
     2401 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2461 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2521 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2581 gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcttaatt
     2641 aagacttcta atataacttc gtatagcata cattatacga agttatggtt ccactgttga
     2701 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     2761 ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2821 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2881 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2941 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3001 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3061 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3121 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3181 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3241 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3301 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3361 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3421 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     3481 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3541 tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc
     3601 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3661 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3721 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3781 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3841 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3901 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3961 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4021 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4081 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4141 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4201 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4261 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
     4321 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4381 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt ccggaatcgt
     4441 cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg cggccatcgt
     4501 gatggctaga gtttagccct cccacacata accagagggc agcaattcac gaatcccaac
     4561 tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat gcagatcgag
     4621 aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat ttccgatcgc
     4681 gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca ccacgagttc
     4741 tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc ggccgtcgct
     4801 agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt tgattgtagc
     4861 cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca tggtggcgct
     4921 agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     4981 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     5041 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     5101 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5161 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     5221 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     5281 ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg ttatttcagg
     5341 ccatggtgct gcgc
//
