LOCUS       sequence-423431-        5690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW2. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5690)
  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 5690)
  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_FBXW2:1-454
FEATURES             Location/Qualifiers
     source          1..5690
                     /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    2753..2874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2764..2783)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2989..3022)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3032..3079
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3111..3905
                     /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(3165..3184)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3775..3794
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4063..4651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4143..4162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4777..4825
                     /note="synthetic polyadenylation signal"
     CDS             complement(4849..5247)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5244..5253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5278..5635)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5292..5427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5444..5635
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5592..5612
                     /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 ccatggagag aaaggacttt
     1261 gagacatggc ttgataacat ttctgttaca tttctttctc tgacggactt gcagaaaaat
     1321 gaaactctgg atcacctgat tagtctgagt ggggcagtcc agctcaggca tctctccaat
     1381 aacctagaga ctctcctcaa gcgggacttc ctcaaactcc ttcccctgga gctcagtttt
     1441 tatttgttaa aatggctcga tcctcagact ttactcacat gctgcctcgt ctctaaacag
     1501 tggaataagg tgataagtgc ctgtacagag gtgtggcaga ctgcatgtaa aaatttgggc
     1561 tggcagatag atgattctgt tcaggacgct ttgcactgga agaaggttta tttgaaggct
     1621 attttgagaa tgaagcaact ggaggaccat gaagcctttg aaacctcgtc attaattgga
     1681 cacagtgcca gagtgtatgc actttactac aaagatggac ttctctgtac agggtcagat
     1741 gacttgtctg caaagctgtg ggatgtgagc acagggcagt gcgtttatgg catccagacc
     1801 cacacttgtg cagcggtgaa gtttgatgaa cagaagcttg tgacaggctc ctttgacaac
     1861 actgtggctt gctgggaatg gagttccgga gccaggaccc agcactttcg ggggcacacg
     1921 ggggcggtat ttagcgtgga ctacaatgat gaactggata tcttggtgag cggctctgca
     1981 gacttcactg tgaaagtatg ggctttatct gctgggacat gcctgaacac actcaccggg
     2041 cacacggaat gggtcaccaa ggtagttttg cagaagtgca aagtcaagtc tctcttgcac
     2101 agtcctggag actacatcct cttaagtgca gacaaatatg agattaagat ttggccaatt
     2161 gggagagaaa tcaactgtaa gtgcttaaag acattgtctg tctctgagga tagaagtatc
     2221 tgcctgcagc caagacttca ttttgatggc aaatacattg tctgtagttc agcacttggt
     2281 ctctaccagt gggactttgc cagttatgat attctcaggg tcatcaagac tcctgagata
     2341 gcaaacttgg ccttgcttgg ctttggagat atctttgccc tgctgtttga caaccgctac
     2401 ctgtacatca tggacttgcg gacagagagc ctgattagtc gctggcctct gccagagtac
     2461 aggaaatcaa agagaggctc aagcttcctg gcaggcgaag catcctggct gaatggactg
     2521 gatgggcaca atgacacggg cttggtcttt gccaccagca tgcctgacca cagtattcac
     2581 ctggtgttgt ggaaggagca cggcggctcg agcggtggga gctccggtgt gagcggctgg
     2641 cggctgttca agaagattag ctaagtttaa acggccgcga ctctagactg caggcatgca
     2701 agctgatttg gctgctgcca ccgctgagca aggccgcttc gagcagacat gataagatac
     2761 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2821 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2881 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc
     2941 aagtaaaacc tctacaaatg tggtaaaatc ttaattaaga cttctaatat aacttcgtat
     3001 agcatacatt atacgaagtt atggttccac tgttgacaat taatcatcgg catagtatat
     3061 cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac
     3121 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3181 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3241 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3301 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3361 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3421 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3481 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3541 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3601 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3661 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3721 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3781 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3841 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3901 tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     3961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4021 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4501 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4621 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     4681 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4741 gatcttcacc tagatccttt tatagtccgg aatcgtcaca caaaaaacca acacacagat
     4801 gtaatgaaaa taaagatatt ttattgcggc catcgtgatg gctagagttt agccctccca
     4861 cacataacca gagggcagca attcacgaat cccaactgcc gtcggctgtc catcactgtc
     4921 cttcactatg gctttgatcc caggatgcag atcgagaagc acctgtcggc accgtccgca
     4981 ggggctcaag atgcccctgt tctcatttcc gatcgcgacg atacaagtca ggttgccagc
     5041 tgccgcagca gcagcagtgc ccagcaccac gagttctgca caaggtcccc cagtaaaatg
     5101 atatacattg acaccagtga agatgcggcc gtcgctagag agagctgcgc tggcgacgct
     5161 gtagtcttca gagatgggga tgctgttgat tgtagccgtt gctctttcaa tgagggtgga
     5221 ttcttcttga gacaaaggct tggccatggt ggcgctagtg tcagaagaat cgagcttttt
     5281 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     5341 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     5401 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     5461 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     5521 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     5581 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcagaaggt
     5641 acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc
//
