LOCUS       sequence-423429-        7511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW10 isoform 2. 
            C-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7511)
  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 7511)
  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_FBXW10:1-1061
FEATURES             Location/Qualifiers
     source          1..7511
                     /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"
     regulatory      2254..2263
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4574..4695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4585..4604)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4810..4843)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4853..4900
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4932..5726
                     /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(4986..5005)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5596..5615
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5884..6472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5964..5983)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6598..6646
                     /note="synthetic polyadenylation signal"
     CDS             complement(6670..7068)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7065..7074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7099..7456)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7113..7248
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7265..7456
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7413..7433
                     /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 ccatggaaaa cctggaatca
     1261 aggctcaaga atgcccccta ttttcgttgt gagaagggaa ccgattccat ccctctatgc
     1321 cggaagtgtg agacgtgtgt cttagcctgg aagatcttct ctaccaaaga gtggttctgc
     1381 aggatcaatg acatatcaca gaggaggttt ctagttggca ttctgaagca gttaaatagc
     1441 ttatatttgt tacactattt ccaaaatatc cttcagacca cacagggaaa ggatttcatc
     1501 tataacaggt cccggatcaa cctcagcaag aaagagggga aagttgtgaa gtcctccttg
     1561 aaccaaatgt tggataaaac agtagaacag aagatgaaag agatcttgta ctggtttgcg
     1621 aacagcaccc agtggaccaa ggcgaattat actctcttac tgctgcagat gtgcaacccc
     1681 aaattactgc tcactgctgc caatgtgatc agagtcctgt ttctgagaga ggagaacaat
     1741 atctcagggc tcaatcaaga catcacagat gtgtgttttt cccctgagaa agaccacagc
     1801 tccaagtctg cgacctcaca agtctattgg acagccaaaa ctcagcacac atcccttcct
     1861 ttgtccaaag ccccagaaaa tgaacacttg cttggggcag catctaaccc tgaggaacca
     1921 tggaggaatt cactccggtg tatatccgaa atgaataggc tgttttctgg aaaaggagac
     1981 ataaccaagc cagggtacga tccctgcaat ctattggttg acctggatga catcagagac
     2041 ctgtcttctg ggttcagcaa atacctagac ttcatccgtt acctgcccat ccacctctcc
     2101 aagtacattc taagtatgct ggggtgtatg aggggatttc ccagacacct cccatcttgg
     2161 cccagttctg gggtttctgg gtccatcttt ttttttccag gaatgctgga tagacacacc
     2221 ctgaacaagt gcgcctctgt gagccagcac tgggccgcca tggctcaaca ggtcaagatg
     2281 gacttgtcag cgcacggctt cattcagaac cagattacct tcttgcaggg gtcctacaca
     2341 agaggaattg atcctaatta tgccaataag gtttctatcc cagttcctaa aatggtagat
     2401 gacgggaaga gcatgcgtgt gaaacatccg aagtggaagc tgagaacgaa gaatgagtac
     2461 aacctgtgga ctgcatacca gaacgaggaa acgcagcagg tcctgataga ggagagaaat
     2521 gttttctgtg ggacctacaa tgttcgcatt ctctctgaca cgtgggatca aaaccgagtc
     2581 atccactatt ccgggggaga tctgatagct gtgtcatcta atcgaaagat ccatcttctg
     2641 gacatcatac aagtgaaagc gatacccgtt gaattccgag gccatgctgg gagtgtccgg
     2701 gccctcttcc tgtgtgagga ggaaaacttt ctcctaagcg ggagctatga cctaagtatc
     2761 agatactggg atctgaaaag tggggtttgc acacgaatct tcggtggtca ccaggggact
     2821 atcacttgca tggacttgtg taagaacagg ctcgtatctg gaggaagaga ttgccaggta
     2881 aaagtatggg atgtagacac agggaagtgc ctgaagacgt ttagacacaa agaccccatc
     2941 ttggccacca ggatcaatga tacctacatt gtgagcagct gtgagcgagg gctggtgaaa
     3001 gtgtggcaca ttgccatggc ccagttggta aagactctca gtggccatga gggagccgtg
     3061 aaatgcctgt tctttgacca gtggcatctc ctctcaggaa gtactgatgg cctggtcatg
     3121 gcctggagca tggtggggaa gtacgagcgc tgcctgatgg ccttcaagca tcccaaggag
     3181 gtgctcgacg tgtccctcct cttcctccgg gtcatcagcg cctgtgcaga tggcaagatc
     3241 cgaatttaca atttcctcaa cgggaactgt atgaaggtgt taaaagccaa tggcagaggt
     3301 gatcctgtgc tgtccttctt tattcagggc aacaggatgg tggtcaacac agagagcaat
     3361 gttctcatgt tccagtttga gcacataaag tggcagtatg ccgtggaaaa aacgaaacaa
     3421 aagaagaata aggagaaaga ggaggaaaaa gaagaaaata gtctcatgga aattctctct
     3481 aagtgtaata ttcaggttca cagcccaaga gagtctgtat ccagtaaaca aactgtgatc
     3541 caagagctcc taccaggcaa acctcccaag tcccgagtac tcctgaagcc ggccaagttc
     3601 tcttcagatg atgtggagaa agcacaaaaa caaggacaat tggaaactcc tggaaaactg
     3661 cccagtcacc caaagaaaaa gtcttggaaa atccctatgt cacctgacca attcctcctg
     3721 actgttagcg ccctgcagca cgcccataat tccggggaat ttgcctatcc ctgtaggccc
     3781 caaacagaaa ttactgatgt ctggggacct tcaatttcat acccaaggaa ggtcttgaat
     3841 ttcaaaggaa aatcaatcca acgtgcagtt gatcggttga gattgagcaa tcctcctata
     3901 gatgtgaaac gaaccagtat tccccttgaa atccagaaac tgcagcccaa cttgaagatc
     3961 tctttgcaca gtcctagagt ccagtccacc ataccccagc ccatgattat ccgctccagg
     4021 ttctctggca gcttaaaggg tggagaccaa gtgaccagtt caattgaaag ggctgtgtgc
     4081 agtacgggtc ccctgaccag tatgcaggtc attaaaccaa accgcatgct agctccacaa
     4141 gtgggcacag ccaccctgtc tcttaagaaa gaacggcctc gcatctatac agcccttgat
     4201 ccttttagag tgaacactga gttcgtgctg ttgaccgtga aggaggagaa ggagcaccag
     4261 gaagccaaga tgaaggaata tcaggccagg gagtccactg gagtggttga tccaggaaaa
     4321 gtcagcaaag ctgcatggat caggaagatc aaaggcctgc ctattgataa tttcacgaag
     4381 caagggaaaa cagcggcccc tgaacttgga caaaatgtat ttatcggctc gagcggtggg
     4441 agctccggtg tgagcggctg gcggctgttc aagaagatta gctaagttta aacggccgcg
     4501 actctagact gcaggcatgc aagctgattt ggctgctgcc accgctgagc aaggccgctt
     4561 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4621 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4681 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4741 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cttaattaag
     4801 acttctaata taacttcgta tagcatacat tatacgaagt tatggttcca ctgttgacaa
     4861 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca
     4921 ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4981 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5041 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5101 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5161 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5221 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5281 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5341 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5401 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5461 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5521 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5581 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5641 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5701 atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg cccaaccggt
     5761 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5821 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5881 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5941 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6001 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6061 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6121 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6181 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6241 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6301 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6361 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6421 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     6481 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6541 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaatcgtcac
     6601 acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg ccatcgtgat
     6661 ggctagagtt tagccctccc acacataacc agagggcagc aattcacgaa tcccaactgc
     6721 cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca gatcgagaag
     6781 cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc cgatcgcgac
     6841 gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca cgagttctgc
     6901 acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc cgtcgctaga
     6961 gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga ttgtagccgt
     7021 tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg tggcgctagt
     7081 gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     7141 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     7201 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     7261 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     7321 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     7381 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg
     7441 gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta tttcaggcca
     7501 tggtgctgcg c
//
