LOCUS       sequence-425875-        5318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human WDHD1 fragment M1 to D330. C-terminal 
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5318)
  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 5318)
  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_WDHD1:1-330
FEATURES             Location/Qualifiers
     source          1..5318
                     /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    2381..2502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2392..2411)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2617..2650)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2660..2707
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2739..3533
                     /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(2793..2812)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3403..3422
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3691..4279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3771..3790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4405..4453
                     /note="synthetic polyadenylation signal"
     CDS             complement(4477..4875)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4872..4881
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4906..5263)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4920..5055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5072..5263
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5220..5240
                     /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 atggctcgag cggtgggagc tccggtgtga gcggctggcg gctgttcaag
     2281 aagattagct aagtttaaac ggccgcgact ctagactgca ggcatgcaag ctgatttggc
     2341 tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat tgatgagttt
     2401 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2461 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2521 cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc
     2581 tacaaatgtg gtaaaatctt aattaagact tctaatataa cttcgtatag catacattat
     2641 acgaagttat ggttccactg ttgacaatta atcatcggca tagtatatcg gcatagtata
     2701 atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc
     2761 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2821 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2881 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     2941 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3001 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3061 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3121 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3181 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3241 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3301 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3361 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3421 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3481 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaaa
     3541 tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc
     3601 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3661 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3721 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3781 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3841 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3901 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3961 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4021 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4081 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     4141 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4201 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4261 cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4321 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4381 gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt aatgaaaata
     4441 aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca cataaccaga
     4501 gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct tcactatggc
     4561 tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat
     4621 gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc
     4681 agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat atacattgac
     4741 accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt agtcttcaga
     4801 gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt cttcttgaga
     4861 caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc aaaagcctag
     4921 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     4981 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     5041 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     5101 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     5161 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     5221 ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac ctaaccaagt
     5281 tcctctttca gaggttattt caggccatgg tgctgcgc
//
