LOCUS       sequence-423383-        5982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW5. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5982)
  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 5982)
  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_FBXW5:1-566
FEATURES             Location/Qualifiers
     source          1..5982
                     /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    3045..3166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3056..3075)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3281..3314)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3324..3371
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3403..4197
                     /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(3457..3476)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4067..4086
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4355..4943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4435..4454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5069..5117
                     /note="synthetic polyadenylation signal"
     CDS             complement(5141..5539)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5536..5545
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5570..5927)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5584..5719
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5736..5927
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5884..5904
                     /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 tggacgaggg
     1261 cggcacgccc ctgctccccg acagcctggt ctaccagatc ttcctgagcc tgggcccggc
     1321 cgacgtgctg gccgccgggc tggtgtgccg ccaatggcag gccgtgtcgc gggacgagtt
     1381 cctgtggagg gagcagttct accgctacta ccaggtggcc cgcgacgtgc cccgacaccc
     1441 agcggccatg tcctggtacg aggagttcca gcggctgtat gacacggtgc cctgcgtgga
     1501 ggtgcagacg ctgcgggaac acacagacca ggtcctgcac ctcagcttct cccattccgg
     1561 gtaccagttc gcgtcctgct ccaaggactg cactgtgaag atctggagca acgacctgac
     1621 catctcgctg ctgcacagcg cggacatgcg gccctacaac tggagctaca cccagttctc
     1681 ccagttcaac aaggacgact cgctactgct ggcctcgggg gtgttcctgg ggccgcacaa
     1741 ctcctcatcc ggcgagattg ctgtcatcag cctagactcc ttcgcgctgc tgtcccgcgt
     1801 gcggaacaag ccctatgacg tgtttggctg ttggctcacc gagaccagcc tcatctcggg
     1861 gaacctgcac cgcatcggag atatcacctc ctgctcggtg ctgtggctca acaatgcctt
     1921 ccaggatgtg gagtcagaga acgtcaacgt ggtgaagcgg ctgttcaaga tccagaacct
     1981 caatgccagc accgtccgca cggtgatggt ggccgactgc agccgcttcg acagccctga
     2041 cctgctgctg gaagccggtg acccggccac gtccccctgc cgcatctttg acctgggcag
     2101 cgacaacgag gaggtggtgg ctggcccggc ccccgcccac gccaaggagg gcttgcggca
     2161 ctttctggac cgcgtgctgg aggggcgggc gcagccacag ctgtcggagc gcatgctaga
     2221 gaccaaggtg gccgagctgc tggcccaggg ccacaccaag ccacccgagc gcagtgccac
     2281 aggcgccaag agcaagtacc tcatcttcac cactggctgc ctcacctact ccccacacca
     2341 gatcggcatc aagcagatcc tgccacacca gatgaccacg gcagggcccg tgctgggtga
     2401 gggccggggc tccgatgcct tcttcgacgc gctggaccac gtcatagaca tacacggaca
     2461 catcatcggc atgggcctgt cgcccgacaa caggtacctg tacgtgaaca gccgcgcctg
     2521 gcccaacggt gcggtggtgg ccgaccccat gcagccgcca ccaatcgcgg aggagattga
     2581 cctgctggtg ttcgacctca agaccatgcg ggaggtgagg cgggctctgc gtgcgcaccg
     2641 cgcctacacg cccaacgacg agtgcttctt catcttcctg gacgtcagca gggacttcgt
     2701 ggccagcggg gcggaggacc ggcacggcta catctgggac cgccactaca acatctgtct
     2761 ggccaggctg cggcacgagg atgtggtcaa ctcagtggtc ttcagtcccc aggagcagga
     2821 gctgctgctc acggccagcg acgacgccac catcaaagcc tggcgctccc cacgcaccat
     2881 gcgcgtcctc caggcacctc gcccacggcc tcgcaccttc ttctcctggc ttgccagcca
     2941 gaggcgctaa tctagaggtc aagcttcggg ggtttaaacc tgcaggcatg caagctgatt
     3001 tggctgctgc caccgctgag caaggccgct tcgagcagac atgataagat acattgatga
     3061 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3121 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3181 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     3241 cctctacaaa tgtggtaaaa tcttaattaa gacttctaat ataacttcgt atagcataca
     3301 ttatacgaag ttatggttcc actgttgaca attaatcatc ggcatagtat atcggcatag
     3361 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga
     3421 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3481 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3541 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3601 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3661 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3721 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3781 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3841 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3901 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     3961 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4021 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4081 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4141 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc
     4201 gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt
     4261 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4321 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4381 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4441 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4501 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4561 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4621 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4681 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4741 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4801 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4861 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4921 cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4981 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5041 cctagatcct tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa
     5101 aataaagata ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac
     5161 cagagggcag caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta
     5221 tggctttgat cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca
     5281 agatgcccct gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag
     5341 cagcagcagt gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat
     5401 tgacaccagt gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt
     5461 cagagatggg gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt
     5521 gagacaaagg cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc
     5581 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     5641 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     5701 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     5761 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     5821 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     5881 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc
     5941 aagttcctct ttcagaggtt atttcaggcc atggtgctgc gc
//
