LOCUS       sequence-423405-        6000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human WDR33 fragment M1 to I572. N-terminal 
            HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6000)
  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 6000)
  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_WDR33:1-572
FEATURES             Location/Qualifiers
     source          1..6000
                     /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    3063..3184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3074..3093)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3299..3332)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3342..3389
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3421..4215
                     /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(3475..3494)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4085..4104
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4373..4961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4453..4472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5087..5135
                     /note="synthetic polyadenylation signal"
     CDS             complement(5159..5557)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5554..5563
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5588..5945)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5602..5737
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5754..5945
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5902..5922
                     /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 tggctacaga
     1261 aattggttct cctcctcgtt ttttccatat gccaaggttc cagcaccagg cacctcgaca
     1321 gctgttttat aagcgacctg attttgcaca acagcaagca atgcaacagc ttacttttga
     1381 tggaaaacga atgagaaaag ctgtgaaccg aaaaaccata gactacaatc catctgtaat
     1441 taagtatttg gagaacagaa tatggcaaag agaccagaga gatatgcggg caattcagcc
     1501 tgatgcaggt tattacaatg atctggtccc acctatagga atgttgaata atcctatgaa
     1561 tgcagtaaca acaaaatttg ttcggacatc aacaaataaa gtaaagtgtc ctgtatttgt
     1621 tgttaggtgg actccagaag gaagacgctt ggtcactgga gcttctagtg gggagtttac
     1681 cctgtggaat ggactcactt tcaattttga aacaatatta caggctcacg acagcccagt
     1741 gagggccatg acgtggtcac ataatgacat gtggatgttg acagcagacc acggaggata
     1801 tgtgaaatat tggcagtcga acatgaacaa cgtcaagatg ttccaggcac ataaggaggc
     1861 gattagagag gccagtttct cacccacgga taataaattt gctacatgct ctgatgacgg
     1921 cactgttaga atctgggact ttcttcgttg ccatgaggaa agaattctcc gagggcatgg
     1981 tgctgatgtg aaatgtgtag actggcatcc aaccaaaggg ttagttgttt caggaagtaa
     2041 agatagtcaa cagccaatca agttctggga tcccaagact gggcagagtc ttgcaacact
     2101 tcatgcccat aaaaacacag taatggaagt gaaattaaac ctcaatggca attggctact
     2161 cacagcatca cgtgatcatc tctgtaaact ttttgatatc agaaacctaa aagaagagct
     2221 tcaagtcttc cgaggtcata agaaagaagc cacagctgtg gcctggcatc ctgttcatga
     2281 aggacttttt gccagtggag ggtctgatgg ttctttgtta ttctggcatg ttggggtaga
     2341 gaaggaagtg ggtgggatgg agatggctca cgaagggatg atctggagtc tggcttggca
     2401 tcctcttggg catattctct gctcaggctc aaatgaccat actagcaaat tctggactcg
     2461 aaaccgacca ggtgataaaa tgcgagatcg atataatcta aaccttttac ctggaatgtc
     2521 tgaagatgga gtagaatatg atgacctcga acctaatagc ctggcagtaa ttccaggaat
     2581 gggaatacca gaacaactaa aattagctat ggaacaagaa cagatgggga aagatgaatc
     2641 aaatgaaatt gaaatgacaa ttccaggttt agattgggga atggaggaag tgatgcaaaa
     2701 ggatcagaaa aaagtacctc agaagaaagt tccttatgca aaacccattc ctgctcagtt
     2761 ccagcaggct tggatgcaaa ataaagttcc aattcctgct ccaaatgagg tgctgaatga
     2821 cagaaaagaa gacattaaat tggaagagaa gaaaaaaaca caagcagaaa ttgagcaaga
     2881 aatggctaca ttacaatata ctaacccaca acttctggag caacttaaaa ttgaaagact
     2941 tgcacagaaa caagttgagc aaatttaatc tagaggtcaa gcttcggggg tttaaacctg
     3001 caggcatgca agctgatttg gctgctgcca ccgctgagca aggccgcttc gagcagacat
     3061 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3121 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3181 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     3241 ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc ttaattaaga cttctaatat
     3301 aacttcgtat agcatacatt atacgaagtt atggttccac tgttgacaat taatcatcgg
     3361 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc
     3421 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3481 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3541 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3601 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3661 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3721 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3781 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3841 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3901 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3961 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4021 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4081 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4141 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4201 gacgagttct tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact
     4261 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4321 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4381 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4441 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4501 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4561 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4621 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4681 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4741 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4801 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4861 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4921 tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt
     4981 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5041 tatcaaaaag gatcttcacc tagatccttt tatagtccgg aatcgtcaca caaaaaacca
     5101 acacacagat gtaatgaaaa taaagatatt ttattgcggc catcgtgatg gctagagttt
     5161 agccctccca cacataacca gagggcagca attcacgaat cccaactgcc gtcggctgtc
     5221 catcactgtc cttcactatg gctttgatcc caggatgcag atcgagaagc acctgtcggc
     5281 accgtccgca ggggctcaag atgcccctgt tctcatttcc gatcgcgacg atacaagtca
     5341 ggttgccagc tgccgcagca gcagcagtgc ccagcaccac gagttctgca caaggtcccc
     5401 cagtaaaatg atatacattg acaccagtga agatgcggcc gtcgctagag agagctgcgc
     5461 tggcgacgct gtagtcttca gagatgggga tgctgttgat tgtagccgtt gctctttcaa
     5521 tgagggtgga ttcttcttga gacaaaggct tggccatggt ggcgctagtg tcagaagaat
     5581 cgagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     5641 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     5701 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     5761 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     5821 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5881 gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc
     5941 ctcagaaggt acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc
//
