LOCUS       sequence-423472-        6044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human WDR33 fragment M1 to I572. C-terminal 
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6044)
  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 6044)
  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_WDR33:1-572
FEATURES             Location/Qualifiers
     source          1..6044
                     /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    3107..3228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3118..3137)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3343..3376)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3386..3433
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3465..4259
                     /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(3519..3538)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4129..4148
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4417..5005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4497..4516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5131..5179
                     /note="synthetic polyadenylation signal"
     CDS             complement(5203..5601)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5598..5607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5632..5989)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5646..5781
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5798..5989
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5946..5966
                     /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 ccatggctac agaaattggt
     1261 tctcctcctc gttttttcca tatgccaagg ttccagcacc aggcacctcg acagctgttt
     1321 tataagcgac ctgattttgc acaacagcaa gcaatgcaac agcttacttt tgatggaaaa
     1381 cgaatgagaa aagctgtgaa ccgaaaaacc atagactaca atccatctgt aattaagtat
     1441 ttggagaaca gaatatggca aagagaccag agagatatgc gggcaattca gcctgatgca
     1501 ggttattaca atgatctggt cccacctata ggaatgttga ataatcctat gaatgcagta
     1561 acaacaaaat ttgttcggac atcaacaaat aaagtaaagt gtcctgtatt tgttgttagg
     1621 tggactccag aaggaagacg cttggtcact ggagcttcta gtggggagtt taccctgtgg
     1681 aatggactca ctttcaattt tgaaacaata ttacaggctc acgacagccc agtgagggcc
     1741 atgacgtggt cacataatga catgtggatg ttgacagcag accacggagg atatgtgaaa
     1801 tattggcagt cgaacatgaa caacgtcaag atgttccagg cacataagga ggcgattaga
     1861 gaggccagtt tctcacccac ggataataaa tttgctacat gctctgatga cggcactgtt
     1921 agaatctggg actttcttcg ttgccatgag gaaagaattc tccgagggca tggtgctgat
     1981 gtgaaatgtg tagactggca tccaaccaaa gggttagttg tttcaggaag taaagatagt
     2041 caacagccaa tcaagttctg ggatcccaag actgggcaga gtcttgcaac acttcatgcc
     2101 cataaaaaca cagtaatgga agtgaaatta aacctcaatg gcaattggct actcacagca
     2161 tcacgtgatc atctctgtaa actttttgat atcagaaacc taaaagaaga gcttcaagtc
     2221 ttccgaggtc ataagaaaga agccacagct gtggcctggc atcctgttca tgaaggactt
     2281 tttgccagtg gagggtctga tggttctttg ttattctggc atgttggggt agagaaggaa
     2341 gtgggtggga tggagatggc tcacgaaggg atgatctgga gtctggcttg gcatcctctt
     2401 gggcatattc tctgctcagg ctcaaatgac catactagca aattctggac tcgaaaccga
     2461 ccaggtgata aaatgcgaga tcgatataat ctaaaccttt tacctggaat gtctgaagat
     2521 ggagtagaat atgatgacct cgaacctaat agcctggcag taattccagg aatgggaata
     2581 ccagaacaac taaaattagc tatggaacaa gaacagatgg ggaaagatga atcaaatgaa
     2641 attgaaatga caattccagg tttagattgg ggaatggagg aagtgatgca aaaggatcag
     2701 aaaaaagtac ctcagaagaa agttccttat gcaaaaccca ttcctgctca gttccagcag
     2761 gcttggatgc aaaataaagt tccaattcct gctccaaatg aggtgctgaa tgacagaaaa
     2821 gaagacatta aattggaaga gaagaaaaaa acacaagcag aaattgagca agaaatggct
     2881 acattacaat atactaaccc acaacttctg gagcaactta aaattgaaag acttgcacag
     2941 aaacaagttg agcaaattgg ctcgagcggt gggagctccg gtgtgagcgg ctggcggctg
     3001 ttcaagaaga ttagctaagt ttaaacggcc gcgactctag actgcaggca tgcaagctga
     3061 tttggctgct gccaccgctg agcaaggccg cttcgagcag acatgataag atacattgat
     3121 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3181 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3241 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa
     3301 aacctctaca aatgtggtaa aatcttaatt aagacttcta atataacttc gtatagcata
     3361 cattatacga agttatggtt ccactgttga caattaatca tcggcatagt atatcggcat
     3421 agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt gaacaagatg
     3481 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3541 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3601 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3661 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3721 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3781 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3841 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3901 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3961 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4021 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4081 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4141 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4201 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgat
     4261 tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg
     4321 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4381 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4441 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4501 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4561 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4621 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4681 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4741 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4801 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4861 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4921 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4981 tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5101 cacctagatc cttttatagt ccggaatcgt cacacaaaaa accaacacac agatgtaatg
     5161 aaaataaaga tattttattg cggccatcgt gatggctaga gtttagccct cccacacata
     5221 accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac tgtccttcac
     5281 tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc cgcaggggct
     5341 caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc
     5401 agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa aatgatatac
     5461 attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga cgctgtagtc
     5521 ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg tggattcttc
     5581 ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa
     5641 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     5701 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     5761 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     5821 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     5881 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     5941 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga aggtacctaa
     6001 ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgc
//
