LOCUS       sequence-423375-        5597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR12. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5597)
  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 5597)
  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_WDR12:1-423
FEATURES             Location/Qualifiers
     source          1..5597
                     /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    2660..2781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2671..2690)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2896..2929)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2939..2986
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3018..3812
                     /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(3072..3091)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3682..3701
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3970..4558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4050..4069)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4684..4732
                     /note="synthetic polyadenylation signal"
     CDS             complement(4756..5154)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5151..5160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5185..5542)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5199..5334
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5351..5542
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5499..5519
                     /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 ccatggctca gctccaaaca
     1261 cgcttctaca ctgataacaa gaaatatgcc gtagatgatg ttcccttctc aatccctgct
     1321 gcctctgaaa ttgccgacct tagtaacatc atcaataaac tactaaagga caaaaatgag
     1381 ttccacaaac atgtggagtt tgatttcctt attaagggcc agtttctgcg aatgcccttg
     1441 gacaaacaca tggaaatgga gaacatctca tcagaagaag ttgtggaaat agaatacgtg
     1501 gagaagtata ctgcacccca gccagagcaa tgcatgttcc atgatgactg gatcagttca
     1561 attaaagggg cagaggaatg gatcttgact ggttcttatg ataagacttc tcggatctgg
     1621 tccttggaag gaaagtcaat aatgacaatt gtgggacata cggatgttgt aaaagatgtg
     1681 gcctgggtga aaaaagatag tttgtcctgc ttattattga gtgcttctat ggatcagact
     1741 attctcttat gggagtggaa tgtagagaga aacaaagtga aagccctaca ctgctgtaga
     1801 ggtcatgctg gaagtgtaga ttctatagct gttgatggct caggaactaa attttgcagt
     1861 ggctcctggg ataagatgct aaagatctgg tctacagtcc ctacagatga agaagatgaa
     1921 atggaggagt ccacaaatcg accaagaaag aaacagaaga cagaacagtt gggactaaca
     1981 aggactccca tagtgaccct ctctggccac atggaggcag tttcctcagt tctgtggtca
     2041 gatgctgaag aaatctgcag tgcatcttgg gaccatacaa ttagagtgtg ggatgttgag
     2101 tctggcagtc ttaagtcaac tttgacagga aataaagtgt ttaattgtat ttcctattct
     2161 ccactttgta aacgtttagc atctggaagc acagataggc atatcagact gtgggatccc
     2221 cgaactaaag atggttcttt ggtgtcgctg tccctaacgt cacatactgg ttgggtgaca
     2281 tcagtaaaat ggtctcctac ccatgaacag cagctgattt caggatcttt agataacatt
     2341 gttaagctgt gggatacaag aagttgtaag gctcctctct atgatctggc tgctcatgaa
     2401 gacaaagttc tgagtgtaga ctggacagac acagggctac ttctgagtgg aggagcagac
     2461 aataaattgt attcctacag atattcacct accacttccc atgttggggc aggctcgagc
     2521 ggtgggagct ccggtgtgag cggctggcgg ctgttcaaga agattagcta agtttaaacg
     2581 gccgcgactc tagactgcag gcatgcaagc tgatttggct gctgccaccg ctgagcaagg
     2641 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2701 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2761 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2821 ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg taaaatctta
     2881 attaagactt ctaatataac ttcgtatagc atacattata cgaagttatg gttccactgt
     2941 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     3001 aacccaggag gcagatcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3061 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3121 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3181 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3241 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3301 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3361 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3421 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3481 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3541 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3601 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3661 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3721 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3781 ccttctatcg ccttcttgac gagttcttct gattcgaaat gaccgaccaa gcgacgccca
     3841 accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3901 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3961 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4021 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4081 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4141 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4201 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4261 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4321 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4381 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4441 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4501 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga
     4561 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4621 gattttggtc atgagattat caaaaaggat cttcacctag atccttttat agtccggaat
     4681 cgtcacacaa aaaaccaaca cacagatgta atgaaaataa agatatttta ttgcggccat
     4741 cgtgatggct agagtttagc cctcccacac ataaccagag ggcagcaatt cacgaatccc
     4801 aactgccgtc ggctgtccat cactgtcctt cactatggct ttgatcccag gatgcagatc
     4861 gagaagcacc tgtcggcacc gtccgcaggg gctcaagatg cccctgttct catttccgat
     4921 cgcgacgata caagtcaggt tgccagctgc cgcagcagca gcagtgccca gcaccacgag
     4981 ttctgcacaa ggtcccccag taaaatgata tacattgaca ccagtgaaga tgcggccgtc
     5041 gctagagaga gctgcgctgg cgacgctgta gtcttcagag atggggatgc tgttgattgt
     5101 agccgttgct ctttcaatga gggtggattc ttcttgagac aaaggcttgg ccatggtggc
     5161 gctagtgtca gaagaatcga gctttttgca aaagcctagg cctccaaaaa agcctcctca
     5221 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     5281 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     5341 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     5401 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     5461 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     5521 cagctggttc tttccgcctc agaaggtacc taaccaagtt cctctttcag aggttatttc
     5581 aggccatggt gctgcgc
//
