LOCUS       sequence-423471-        5426 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR31 isoform 2. 
            C-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5426)
  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 5426)
  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_WDR31:1-367
FEATURES             Location/Qualifiers
     source          1..5426
                     /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    2489..2610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2500..2519)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2725..2758)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2768..2815
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2847..3641
                     /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(2901..2920)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3511..3530
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3799..4387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3879..3898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4513..4561
                     /note="synthetic polyadenylation signal"
     CDS             complement(4585..4983)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4980..4989
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5014..5371)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5028..5163
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5180..5371
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5328..5348
                     /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 ccatgctgct actcaggtgc
     1261 caactgaaac aagctcctcc acagaaggtt tcgtttaggt tttgtgtcgt gatggggaaa
     1321 cagcaaagca aactcaaaca cagcacttat aaatacgggc ctgatgaaat tatagaagag
     1381 agaattcaaa ctaaagcttt tcaagagtat agcccagctc acatggatac cgtctctgtc
     1441 gtggctgctt tgaactcaga cctttgtgtc tctggaggga aagataagac agttgtggcc
     1501 tataattgga aaactggaaa tgtggtgaaa aggttcaaag gacatgaaca tgagatcacc
     1561 aaggtagcct gtattcccaa atccagccag ttcttcagtg cctctcgtga caggatggtc
     1621 atgatgtggg acttgcacgg ttcctcacaa ccaaggcagc aattgtgtgg ccatgccatg
     1681 gtggtcaccg gattggctgt gagtccagac tcatcacagc tgtgcactgg ctctcgggac
     1741 aacaccctgc ttctgtggga tgtggtgaca ggacagagtg tggaaagagc atctgtctcc
     1801 aggaacgtgg tcactcacct gtgctgggtc cccagagaac catacatact acagacctct
     1861 gaagataaaa ccctcagatt atgggacagt cgggggctgc aggtagctca tatgtttcct
     1921 gcaaagcagc acattcagac ctactgtgaa gtcagtgtgg atggacacaa gtgtatctcc
     1981 tgcagcaatg gctttggagg agaaggctgt gaagccacgt tgtgggacct aagacagact
     2041 cgaaacagaa tatgtgagta taaggggcat ttccagactg tcgcatcctg cgtctttcta
     2101 ccaagagcat tggccttgat gcctttaatt gctacctcat cacatgattg caaggtgaag
     2161 atttggaacc aagatactgg agcctgcctt ttcaccttgt ctctggatgg atcaggaccc
     2221 ttgacttctc tggctgttgg tgacgccatc tccttattgt gtgcaagttt taacagagga
     2281 attcacttac tcagaatgga ccacagccaa gggctggaac tgcaggaagt ggcagcattc
     2341 ggctcgagcg gtgggagctc cggtgtgagc ggctggcggc tgttcaagaa gattagctaa
     2401 gtttaaacgg ccgcgactct agactgcagg catgcaagct gatttggctg ctgccaccgc
     2461 tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     2521 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2581 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2641 caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt
     2701 aaaatcttaa ttaagacttc taatataact tcgtatagca tacattatac gaagttatgg
     2761 ttccactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     2821 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     2881 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2941 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3001 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3061 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3121 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3181 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3241 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3301 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3361 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3421 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3481 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3541 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3601 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     3661 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3721 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3781 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3841 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3901 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3961 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4021 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4081 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4141 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4201 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     4261 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4321 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4381 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4441 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     4501 gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat
     4561 tgcggccatc gtgatggcta gagtttagcc ctcccacaca taaccagagg gcagcaattc
     4621 acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt tgatcccagg
     4681 atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc
     4741 atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag cagtgcccag
     4801 caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac cagtgaagat
     4861 gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga tggggatgct
     4921 gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca aaggcttggc
     4981 catggtggcg ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc ctccaaaaaa
     5041 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     5101 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     5161 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     5221 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     5281 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     5341 cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc ctctttcaga
     5401 ggttatttca ggccatggtg ctgcgc
//
