LOCUS       sequence-425874-        7715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDHD1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7715)
  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 7715)
  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_WDHD1:1-1129
FEATURES             Location/Qualifiers
     source          1..7715
                     /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    4778..4899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4789..4808)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5014..5047)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5057..5104
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5136..5930
                     /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(5190..5209)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5800..5819
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6088..6676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6168..6187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6802..6850
                     /note="synthetic polyadenylation signal"
     CDS             complement(6874..7272)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7269..7278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7303..7660)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7317..7452
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7469..7660
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7617..7637
                     /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 ccatgcctgc cacacggaag
     1261 ccaatgagat atgggcatac agagggacac acggaggtct gttttgatga ttctgggagt
     1321 tttattgtga cttgtggaag tgatggtgat gtgaggattt gggaagactt ggatgatgat
     1381 gatcctaagt tcattaatgt tggagaaaag gcatattcat gtgctttgaa gagtggaaaa
     1441 ctggtcactg cagtttctaa taatactatt caagtccaca catttcctga aggagttcca
     1501 gatggtatat tgactcgctt cactacaaat gcaaaccatg tggtctttaa tggggatggt
     1561 actaaaattg ctgctggatc tagtgatttt ctagtcaaaa ttgtggatgt gatggatagc
     1621 agccaacaga aaacatttcg aggacatgat gcccctgttt taagtctttc ctttgatcct
     1681 aaggacatct ttctggcatc agctagttgt gatggatctg tcagagtgtg gcaaatttca
     1741 gatcagacat gtgctattag ttggccactg ctacaaaaat gcaacgatgt gataaatgca
     1801 aaatcaatct gcagacttgc ttggcagcca aaaagtggga agttactggc aattcctgtg
     1861 gaaaaatctg ttaagctata tagaagagaa tcttggagtc atcaatttga tctttcagat
     1921 aatttcatct ctcagaccct caatatagta acctggtctc cctgtgggca atatttagct
     1981 gcaggtagta ttaatggtct aatcatagtt tggaatgtgg aaaccaaaga ctgcatggaa
     2041 agggtgaaac atgagaaagg ttatgcaatt tgtggtctgg catggcatcc tacttgtggt
     2101 cgaatatcgt atactgatgc ggaaggaaat ctagggcttc tagagaatgt ttgtgacccc
     2161 agtggaaaga catcaagcag taaggtatct agcagagtgg aaaaggatta taatgatctt
     2221 tttgatggag atgatatgag taatgctggt gattttctaa atgacaatgc agttgagatc
     2281 ccttcttttt caaaagggat tataaatgat gatgaggatg atgaagacct catgatggct
     2341 tcaggtcgtc ctagacagcg aagtcacatc ctagaagatg atgaaaactc agttgatatt
     2401 tcaatgctaa aaactggttc tagtcttctc aaagaggagg aggaagatgg tcaagaaggc
     2461 agcattcaca atctaccact tgtaacatcc caaaggccat tttatgatgg acccatgcca
     2521 actccccggc aaaagccatt tcagtcaggt tctacaccgt tgcatctcac tcacagattc
     2581 atggtgtgga actctattgg aattattcgc tgctataatg atgagcaaga caatgccata
     2641 gatgtggagt tccatgatac ctccatacac catgcaacac acttatcaaa cactttgaat
     2701 tatacaatag cagatctttc ccacgaagct attttgttgg catgtgaaag cactgatgaa
     2761 ctagcaagca agcttcactg cctgcacttt agttcttggg attcaagcaa agagtggata
     2821 atagacttgc ctcagaatga ggatattgaa gccatatgtc tcggtcaagg atgggctgct
     2881 gccgctacta gtgccctgct tcttcgattg tttactattg gaggggttca aaaagaggta
     2941 ttcagccttg ctggacctgt ggtgtcaatg gcaggacatg gagaacagct tttcattgtt
     3001 tatcacagag gtacaggatt tgatggggat cagtgccttg gagttcaact gctagagctg
     3061 gggaaaaaga aaaaacaaat tttgcatggt gaccctcttc ctcttacaag gaaatcctac
     3121 cttgcatgga ttgggttttc agctgaaggt accccttgtt acgtggattc agaaggaatt
     3181 gttcgaatgc ttaacagagg acttggtaat acgtggactc ctatatgtaa tacaagagag
     3241 cactgcaaag gaaaatctga tcactactgg gtggttggta tccatgaaaa tccccagcaa
     3301 ctaaggtgca ttccttgtaa aggttctcgg tttcccccaa cccttccacg ccctgctgtt
     3361 gctatattat cctttaagct tccttactgt cagattgcaa cagagaaagg acaaatggag
     3421 gagcaatttt ggcgttcagt tatatttcac aaccaccttg attatttagc taaaaatggt
     3481 tatgaatatg aagagagcac taaaaatcaa gcaacaaaag agcaacagga acttttaatg
     3541 aaaatgcttg cgctttcttg taaactggag cgagaattcc gttgtgtgga acttgctgat
     3601 ctaatgactc aaaatgctgt gaatttagcc attaaatatg cttctcgctc tcggaaatta
     3661 atactggctc aaaaactaag tgaactggct gtagagaagg cagccgaatt gacagcaacc
     3721 caggtggaag aggaagaaga agaagaagat ttcagaaaaa agctgaatgc tggttacagc
     3781 aatactgcta cagagtggag ccaaccaagg ttcagaaatc aagttgaaga agatgctgag
     3841 gacagtggag aagctgatga tgaagaaaaa ccagaaatac ataagcctgg acagaactcg
     3901 ttttccaaaa gtacaaattc ctctgatgtt tcagctaagt caggtgcagt tacctttagc
     3961 agccaaggac gagtaaatcc ctttaaggta tcagccagtt ccaaagaacc agccatgtca
     4021 atgaattcag cacgttcaac taatatttta gacaatatgg gcaaatcatc caagaaatcc
     4081 actgcactta gtcgaactac aaataatgaa aagtctccca ttataaagcc tctgattcca
     4141 aagccgaagc ctaagcaggc atctgcagca tcctatttcc agaaaagaaa ttctcaaact
     4201 aataaaactg aggaagtgaa agaagaaaat cttaaaaatg tattatctga aaccccagct
     4261 atatgtcctc ctcaaaacac tgaaaaccaa aggccaaaga ccgggttcca gatgtggtta
     4321 gaagaaaata gaagtaatat tttgtctgac aatcctgact tttcagatga agcagacata
     4381 ataaaagaag gaatgattcg atttagagta ttgtcaactg aagaaagaaa ggtgtgggct
     4441 aacaaagcca aaggagaaac ggcaagtgaa ggaactgaag caaagaagcg aaaacgtgtg
     4501 gttgatgaaa gtgatgaaac agaaaaccag gaagaaaaag caaaagagaa cctgaatttg
     4561 tctaaaaagc agaaaccttt agatttttct acaaatcaga aactatcagc ttttgcattt
     4621 aagcaggagg gctcgagcgg tgggagctcc ggtgtgagcg gctggcggct gttcaagaag
     4681 attagctaag tttaaacggc cgcgactcta gactgcaggc atgcaagctg atttggctgc
     4741 tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     4801 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     4861 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     4921 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac
     4981 aaatgtggta aaatcttaat taagacttct aatataactt cgtatagcat acattatacg
     5041 aagttatggt tccactgttg acaattaatc atcggcatag tatatcggca tagtataata
     5101 cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg
     5161 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5221 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5281 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5341 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5401 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5461 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5521 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5581 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5641 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5701 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5761 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5821 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5881 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaaatga
     5941 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6001 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6061 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6541 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6661 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6721 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6781 ccttttatag tccggaatcg tcacacaaaa aaccaacaca cagatgtaat gaaaataaag
     6841 atattttatt gcggccatcg tgatggctag agtttagccc tcccacacat aaccagaggg
     6901 cagcaattca cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt
     6961 gatcccagga tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc
     7021 cctgttctca tttccgatcg cgacgataca agtcaggttg ccagctgccg cagcagcagc
     7081 agtgcccagc accacgagtt ctgcacaagg tcccccagta aaatgatata cattgacacc
     7141 agtgaagatg cggccgtcgc tagagagagc tgcgctggcg acgctgtagt cttcagagat
     7201 ggggatgctg ttgattgtag ccgttgctct ttcaatgagg gtggattctt cttgagacaa
     7261 aggcttggcc atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc
     7321 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     7381 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     7441 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat
     7501 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
     7561 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc
     7621 taactgacac acattccaca gctggttctt tccgcctcag aaggtaccta accaagttcc
     7681 tctttcagag gttatttcag gccatggtgc tgcgc
//
