LOCUS       sequence-437049-        7671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDHD1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7671)
  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 7671)
  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_WDHD1:1-1129
FEATURES             Location/Qualifiers
     source          1..7671
                     /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    4734..4855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4745..4764)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4970..5003)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5013..5060
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5092..5886
                     /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(5146..5165)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5756..5775
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6044..6632)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6124..6143)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6758..6806
                     /note="synthetic polyadenylation signal"
     CDS             complement(6830..7228)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7225..7234
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7259..7616)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7273..7408
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7425..7616
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7573..7593
                     /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 tgcctgccac
     1261 acggaagcca atgagatatg ggcatacaga gggacacacg gaggtctgtt ttgatgattc
     1321 tgggagtttt attgtgactt gtggaagtga tggtgatgtg aggatttggg aagacttgga
     1381 tgatgatgat cctaagttca ttaatgttgg agaaaaggca tattcatgtg ctttgaagag
     1441 tggaaaactg gtcactgcag tttctaataa tactattcaa gtccacacat ttcctgaagg
     1501 agttccagat ggtatattga ctcgcttcac tacaaatgca aaccatgtgg tctttaatgg
     1561 ggatggtact aaaattgctg ctggatctag tgattttcta gtcaaaattg tggatgtgat
     1621 ggatagcagc caacagaaaa catttcgagg acatgatgcc cctgttttaa gtctttcctt
     1681 tgatcctaag gacatctttc tggcatcagc tagttgtgat ggatctgtca gagtgtggca
     1741 aatttcagat cagacatgtg ctattagttg gccactgcta caaaaatgca acgatgtgat
     1801 aaatgcaaaa tcaatctgca gacttgcttg gcagccaaaa agtgggaagt tactggcaat
     1861 tcctgtggaa aaatctgtta agctatatag aagagaatct tggagtcatc aatttgatct
     1921 ttcagataat ttcatctctc agaccctcaa tatagtaacc tggtctccct gtgggcaata
     1981 tttagctgca ggtagtatta atggtctaat catagtttgg aatgtggaaa ccaaagactg
     2041 catggaaagg gtgaaacatg agaaaggtta tgcaatttgt ggtctggcat ggcatcctac
     2101 ttgtggtcga atatcgtata ctgatgcgga aggaaatcta gggcttctag agaatgtttg
     2161 tgaccccagt ggaaagacat caagcagtaa ggtatctagc agagtggaaa aggattataa
     2221 tgatcttttt gatggagatg atatgagtaa tgctggtgat tttctaaatg acaatgcagt
     2281 tgagatccct tctttttcaa aagggattat aaatgatgat gaggatgatg aagacctcat
     2341 gatggcttca ggtcgtccta gacagcgaag tcacatccta gaagatgatg aaaactcagt
     2401 tgatatttca atgctaaaaa ctggttctag tcttctcaaa gaggaggagg aagatggtca
     2461 agaaggcagc attcacaatc taccacttgt aacatcccaa aggccatttt atgatggacc
     2521 catgccaact ccccggcaaa agccatttca gtcaggttct acaccgttgc atctcactca
     2581 cagattcatg gtgtggaact ctattggaat tattcgctgc tataatgatg agcaagacaa
     2641 tgccatagat gtggagttcc atgatacctc catacaccat gcaacacact tatcaaacac
     2701 tttgaattat acaatagcag atctttccca cgaagctatt ttgttggcat gtgaaagcac
     2761 tgatgaacta gcaagcaagc ttcactgcct gcactttagt tcttgggatt caagcaaaga
     2821 gtggataata gacttgcctc agaatgagga tattgaagcc atatgtctcg gtcaaggatg
     2881 ggctgctgcc gctactagtg ccctgcttct tcgattgttt actattggag gggttcaaaa
     2941 agaggtattc agccttgctg gacctgtggt gtcaatggca ggacatggag aacagctttt
     3001 cattgtttat cacagaggta caggatttga tggggatcag tgccttggag ttcaactgct
     3061 agagctgggg aaaaagaaaa aacaaatttt gcatggtgac cctcttcctc ttacaaggaa
     3121 atcctacctt gcatggattg ggttttcagc tgaaggtacc ccttgttacg tggattcaga
     3181 aggaattgtt cgaatgctta acagaggact tggtaatacg tggactccta tatgtaatac
     3241 aagagagcac tgcaaaggaa aatctgatca ctactgggtg gttggtatcc atgaaaatcc
     3301 ccagcaacta aggtgcattc cttgtaaagg ttctcggttt cccccaaccc ttccacgccc
     3361 tgctgttgct atattatcct ttaagcttcc ttactgtcag attgcaacag agaaaggaca
     3421 aatggaggag caattttggc gttcagttat atttcacaac caccttgatt atttagctaa
     3481 aaatggttat gaatatgaag agagcactaa aaatcaagca acaaaagagc aacaggaact
     3541 tttaatgaaa atgcttgcgc tttcttgtaa actggagcga gaattccgtt gtgtggaact
     3601 tgctgatcta atgactcaaa atgctgtgaa tttagccatt aaatatgctt ctcgctctcg
     3661 gaaattaata ctggctcaaa aactaagtga actggctgta gagaaggcag ccgaattgac
     3721 agcaacccag gtggaagagg aagaagaaga agaagatttc agaaaaaagc tgaatgctgg
     3781 ttacagcaat actgctacag agtggagcca accaaggttc agaaatcaag ttgaagaaga
     3841 tgctgaggac agtggagaag ctgatgatga agaaaaacca gaaatacata agcctggaca
     3901 gaactcgttt tccaaaagta caaattcctc tgatgtttca gctaagtcag gtgcagttac
     3961 ctttagcagc caaggacgag taaatccctt taaggtatca gccagttcca aagaaccagc
     4021 catgtcaatg aattcagcac gttcaactaa tattttagac aatatgggca aatcatccaa
     4081 gaaatccact gcacttagtc gaactacaaa taatgaaaag tctcccatta taaagcctct
     4141 gattccaaag ccgaagccta agcaggcatc tgcagcatcc tatttccaga aaagaaattc
     4201 tcaaactaat aaaactgagg aagtgaaaga agaaaatctt aaaaatgtat tatctgaaac
     4261 cccagctata tgtcctcctc aaaacactga aaaccaaagg ccaaagaccg ggttccagat
     4321 gtggttagaa gaaaatagaa gtaatatttt gtctgacaat cctgactttt cagatgaagc
     4381 agacataata aaagaaggaa tgattcgatt tagagtattg tcaactgaag aaagaaaggt
     4441 gtgggctaac aaagccaaag gagaaacggc aagtgaagga actgaagcaa agaagcgaaa
     4501 acgtgtggtt gatgaaagtg atgaaacaga aaaccaggaa gaaaaagcaa aagagaacct
     4561 gaatttgtct aaaaagcaga aacctttaga tttttctaca aatcagaaac tatcagcttt
     4621 tgcatttaag caggagtaat ctagaggtca agcttcgggg gtttaaacct gcaggcatgc
     4681 aagctgattt ggctgctgcc accgctgagc aaggccgctt cgagcagaca tgataagata
     4741 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     4801 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     4861 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg tttttttaag
     4921 caagtaaaac ctctacaaat gtggtaaaat cttaattaag acttctaata taacttcgta
     4981 tagcatacat tatacgaagt tatggttcca ctgttgacaa ttaatcatcg gcatagtata
     5041 tcggcatagt ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa
     5101 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5161 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5221 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5281 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5341 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5401 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5461 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5521 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5581 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5641 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5701 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5761 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5821 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5881 ttctgattcg aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg
     5941 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6001 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6061 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6121 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6181 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6241 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6301 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6361 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6421 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6481 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6541 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6601 agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt
     6661 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6721 ggatcttcac ctagatcctt ttatagtccg gaatcgtcac acaaaaaacc aacacacaga
     6781 tgtaatgaaa ataaagatat tttattgcgg ccatcgtgat ggctagagtt tagccctccc
     6841 acacataacc agagggcagc aattcacgaa tcccaactgc cgtcggctgt ccatcactgt
     6901 ccttcactat ggctttgatc ccaggatgca gatcgagaag cacctgtcgg caccgtccgc
     6961 aggggctcaa gatgcccctg ttctcatttc cgatcgcgac gatacaagtc aggttgccag
     7021 ctgccgcagc agcagcagtg cccagcacca cgagttctgc acaaggtccc ccagtaaaat
     7081 gatatacatt gacaccagtg aagatgcggc cgtcgctaga gagagctgcg ctggcgacgc
     7141 tgtagtcttc agagatgggg atgctgttga ttgtagccgt tgctctttca atgagggtgg
     7201 attcttcttg agacaaaggc ttggccatgg tggcgctagt gtcagaagaa tcgagctttt
     7261 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     7321 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     7381 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     7441 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     7501 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     7561 gggactttcc acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg
     7621 tacctaacca agttcctctt tcagaggtta tttcaggcca tggtgctgcg c
//
