LOCUS       sequence-425883-        6290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR70. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  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 6290)
  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_WDR70:1-654
FEATURES             Location/Qualifiers
     source          1..6290
                     /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    3353..3474
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3364..3383)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3589..3622)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3632..3679
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3711..4505
                     /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(3765..3784)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4375..4394
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4663..5251)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4743..4762)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5377..5425
                     /note="synthetic polyadenylation signal"
     CDS             complement(5449..5847)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5844..5853
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5878..6235)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5892..6027
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6044..6235
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6192..6212
                     /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 ccatggagcg ctctgggccc
     1261 agcgaagtga caggctcaga cgcgtcggga ccggacccgc agcttgcggt caccatgggc
     1321 ttcacggggt tcggtaaaaa agctcgcaca tttgacttgg aagcaatgtt tgaacaaact
     1381 cgaaggacag ctgtggaaag aagtcgcaaa acactggaag caagagaaaa agaggaagaa
     1441 atgaacagag agaaagaatt aagaagacaa aatgaagata ttgagccaac atcctcaaga
     1501 tcaaatgtgg tcagagattg ctccaaatca tcttccaggg atacgagcag cagtgaaagt
     1561 gaacagagtt ctgactcttc tgatgatgag ttaattggcc ctcctttacc ccctaaaatg
     1621 gtaggaaaac cagttaattt tatggaggaa gatatcctcg gtcctttacc tccacctctt
     1681 aatgaagaag aagaagaagc agaggaagaa gaagaggaag aggaggaaga ggaaaatcct
     1741 gttcacaaga ttcctgactc gcatgagata acgctgaagc atggcactaa aacagtgtct
     1801 gctttgggtc tggatccctc aggtgcccgt ttggtgacag gaggatatga ctatgatgtt
     1861 aagttttggg attttgctgg aatggatgct tcttttaagg catttcgatc ccttcagccc
     1921 tgtgagtgcc atcagatcaa gtcattacag tatagtaaca caggagacat gattcttgtt
     1981 gtatctggaa gctctcaggc caaggtgatt gacagagatg gttttgaagt aatggaatgt
     2041 ataaaaggag accagtatat tgtggacatg gccaacacca agggtcatac agcaatgctt
     2101 catactggct catggcatcc caaaataaag ggagaattta tgacttgctc aaatgatgcg
     2161 actgtgagga cgtgggaagt tgaaaatcca aagaagcaaa aaagtgtgtt taaaccacgg
     2221 acgatgcaag gcaaaaaagt cattcccact acgtgcacat atagtagaga tggaaacctc
     2281 atagcagctg cctgccagaa tggaagcata cagatctggg accgaaattt gactgttcat
     2341 cctaagttcc actataaaca ggctcatgac tcgggcacag acacttcttg cgtgactttt
     2401 tcctatgatg gtaatgtcct tgcctctcgt ggaggtgacg attcattaaa attatgggac
     2461 atccgacaat ttaataaacc acttttttca gcctcgggtc ttcccaccat gttcccaatg
     2521 actgactgct gtttcagtcc agatgataag ctcatagtca ctggtacatc tattcaaaga
     2581 ggatgtggca gcggcaaact tgttttcttt gagcgtagga ctttccaaag ggtgtatgaa
     2641 atagacatca cagatgcgag tgttgttcgc tgcctgtggc atccaaagct gaaccagatc
     2701 atggttggaa ctggaaatgg attggctaaa gtctattacg accccaacaa gagtcagagg
     2761 ggagcaaaat tatgtgtggt taaaacccag cggaaggcaa aacaagctga gactctaact
     2821 caggactaca tcatcacccc tcatgccttg cctatgttcc gtgagccccg ccaacggagt
     2881 acaaggaaac agctggagaa ggacagactg gatcccctga agtcgcataa acctgaacct
     2941 cctgtagcag gcccaggtcg tggtggccga gttggaaccc acgggggcac tctctcttcc
     3001 tatattgtga agaacattgc tttggacaag accgatgaca gtaatcctcg ggaagccatt
     3061 ttgcgtcatg ccaaagcagc agaagacagc ccatattggg tttctccagc atattccaag
     3121 actcagccca aaaccatgtt tgcccaagtt gaatctgatg atgaggaagc aaagaatgag
     3181 ccagaatgga aaaaacgtaa aattggctcg agcggtggga gctccggtgt gagcggctgg
     3241 cggctgttca agaagattag ctaagtttaa acggccgcga ctctagactg caggcatgca
     3301 agctgatttg gctgctgcca ccgctgagca aggccgcttc gagcagacat gataagatac
     3361 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3421 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3481 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc
     3541 aagtaaaacc tctacaaatg tggtaaaatc ttaattaaga cttctaatat aacttcgtat
     3601 agcatacatt atacgaagtt atggttccac tgttgacaat taatcatcgg catagtatat
     3661 cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac
     3721 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3781 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3841 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3901 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3961 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4021 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4081 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4141 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4201 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4261 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4321 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4381 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4441 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4501 tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     4561 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4621 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4681 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4741 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4801 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4861 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4921 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4981 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5041 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5101 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5161 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5221 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     5281 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5341 gatcttcacc tagatccttt tatagtccgg aatcgtcaca caaaaaacca acacacagat
     5401 gtaatgaaaa taaagatatt ttattgcggc catcgtgatg gctagagttt agccctccca
     5461 cacataacca gagggcagca attcacgaat cccaactgcc gtcggctgtc catcactgtc
     5521 cttcactatg gctttgatcc caggatgcag atcgagaagc acctgtcggc accgtccgca
     5581 ggggctcaag atgcccctgt tctcatttcc gatcgcgacg atacaagtca ggttgccagc
     5641 tgccgcagca gcagcagtgc ccagcaccac gagttctgca caaggtcccc cagtaaaatg
     5701 atatacattg acaccagtga agatgcggcc gtcgctagag agagctgcgc tggcgacgct
     5761 gtagtcttca gagatgggga tgctgttgat tgtagccgtt gctctttcaa tgagggtgga
     5821 ttcttcttga gacaaaggct tggccatggt ggcgctagtg tcagaagaat cgagcttttt
     5881 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     5941 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     6001 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     6061 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     6121 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     6181 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcagaaggt
     6241 acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc
//
