LOCUS       sequence-423404-        5382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR31 isoform 2. 
            N-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5382)
  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 5382)
  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_WDR31:1-367
FEATURES             Location/Qualifiers
     source          1..5382
                     /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    2445..2566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2456..2475)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2681..2714)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2724..2771
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2803..3597
                     /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(2857..2876)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3467..3486
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3755..4343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3835..3854)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4469..4517
                     /note="synthetic polyadenylation signal"
     CDS             complement(4541..4939)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4936..4945
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4970..5327)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4984..5119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5136..5327
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5284..5304
                     /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 tgctgctact
     1261 caggtgccaa ctgaaacaag ctcctccaca gaaggtttcg tttaggtttt gtgtcgtgat
     1321 ggggaaacag caaagcaaac tcaaacacag cacttataaa tacgggcctg atgaaattat
     1381 agaagagaga attcaaacta aagcttttca agagtatagc ccagctcaca tggataccgt
     1441 ctctgtcgtg gctgctttga actcagacct ttgtgtctct ggagggaaag ataagacagt
     1501 tgtggcctat aattggaaaa ctggaaatgt ggtgaaaagg ttcaaaggac atgaacatga
     1561 gatcaccaag gtagcctgta ttcccaaatc cagccagttc ttcagtgcct ctcgtgacag
     1621 gatggtcatg atgtgggact tgcacggttc ctcacaacca aggcagcaat tgtgtggcca
     1681 tgccatggtg gtcaccggat tggctgtgag tccagactca tcacagctgt gcactggctc
     1741 tcgggacaac accctgcttc tgtgggatgt ggtgacagga cagagtgtgg aaagagcatc
     1801 tgtctccagg aacgtggtca ctcacctgtg ctgggtcccc agagaaccat acatactaca
     1861 gacctctgaa gataaaaccc tcagattatg ggacagtcgg gggctgcagg tagctcatat
     1921 gtttcctgca aagcagcaca ttcagaccta ctgtgaagtc agtgtggatg gacacaagtg
     1981 tatctcctgc agcaatggct ttggaggaga aggctgtgaa gccacgttgt gggacctaag
     2041 acagactcga aacagaatat gtgagtataa ggggcatttc cagactgtcg catcctgcgt
     2101 ctttctacca agagcattgg ccttgatgcc tttaattgct acctcatcac atgattgcaa
     2161 ggtgaagatt tggaaccaag atactggagc ctgccttttc accttgtctc tggatggatc
     2221 aggacccttg acttctctgg ctgttggtga cgccatctcc ttattgtgtg caagttttaa
     2281 cagaggaatt cacttactca gaatggacca cagccaaggg ctggaactgc aggaagtggc
     2341 agcattctaa tctagaggtc aagcttcggg ggtttaaacc tgcaggcatg caagctgatt
     2401 tggctgctgc caccgctgag caaggccgct tcgagcagac atgataagat acattgatga
     2461 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2521 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2581 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     2641 cctctacaaa tgtggtaaaa tcttaattaa gacttctaat ataacttcgt atagcataca
     2701 ttatacgaag ttatggttcc actgttgaca attaatcatc ggcatagtat atcggcatag
     2761 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga
     2821 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     2881 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     2941 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3001 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3061 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3121 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3181 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3241 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3301 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     3361 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3421 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3481 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3541 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc
     3601 gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt
     3661 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3721 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3781 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3841 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3901 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3961 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4021 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4081 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4141 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4201 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4261 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4321 cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4381 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4441 cctagatcct tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa
     4501 aataaagata ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac
     4561 cagagggcag caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta
     4621 tggctttgat cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca
     4681 agatgcccct gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag
     4741 cagcagcagt gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat
     4801 tgacaccagt gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt
     4861 cagagatggg gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt
     4921 gagacaaagg cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc
     4981 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     5041 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     5101 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     5161 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     5221 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     5281 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc
     5341 aagttcctct ttcagaggtt atttcaggcc atggtgctgc gc
//
