LOCUS       sequence-425988-        6449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW7. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6449)
  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 6449)
  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_FBXW7:1-707
FEATURES             Location/Qualifiers
     source          1..6449
                     /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    3512..3633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3523..3542)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3748..3781)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3791..3838
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3870..4664
                     /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(3924..3943)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4534..4553
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4822..5410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4902..4921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5536..5584
                     /note="synthetic polyadenylation signal"
     CDS             complement(5608..6006)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6003..6012
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6037..6394)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6051..6186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6203..6394
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6351..6371
                     /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 ccatgaatca ggaactgctc
     1261 tctgtgggca gcaaaagacg acgaactgga ggctctctga gaggtaaccc ttcctcaagc
     1321 caggtagatg aagaacagat gaatcgtgtg gtagaggagg aacagcaaca gcaactcaga
     1381 caacaagagg aggagcacac tgcaaggaat ggtgaagttg ttggagtaga acctagacct
     1441 ggaggccaaa atgattccca gcaaggacag ttggaagaaa acaataatag atttatttcg
     1501 gtagatgagg actcctcagg aaaccaagaa gaacaagagg aagatgaaga acatgctggt
     1561 gaacaagatg aggaggatga ggaggaggag gagatggacc aggagagtga cgattttgat
     1621 cagtctgatg atagtagcag agaagatgaa catacacata ctaacagtgt cacgaactcc
     1681 agtagtattg tggacctgcc cgttcaccaa ctctcctccc cattctatac aaaaacaaca
     1741 aaaatgaaaa gaaagttgga ccatggttct gaggtccgct ctttttcttt gggaaagaaa
     1801 ccatgcaaag tctcagaata tacaagtacc actgggcttg taccatgttc agcaacacca
     1861 acaacttttg gggacctcag agcagccaat ggccaagggc aacaacgacg ccgaattaca
     1921 tctgtccagc cacctacagg cctccaggaa tggctaaaaa tgtttcagag ctggagtgga
     1981 ccagagaaat tgcttgcttt agatgaactc attgatagtt gtgaaccaac acaagtaaaa
     2041 catatgatgc aagtgataga accccagttt caacgagact tcatttcatt gctccctaaa
     2101 gagttggcac tctatgtgct ttcattcctg gaacccaaag acctgctaca agcagctcag
     2161 acatgtcgct actggagaat tttggctgaa gacaaccttc tctggagaga gaaatgcaaa
     2221 gaagagggga ttgatgaacc attgcacatc aagagaagaa aagtaataaa accaggtttc
     2281 atacacagtc catggaaaag tgcatacatc agacagcaca gaattgatac taactggagg
     2341 cgaggagaac tcaaatctcc taaggtgctg aaaggacatg atgatcatgt gatcacatgc
     2401 ttacagtttt gtggtaaccg aatagttagt ggttctgatg acaacacttt aaaagtttgg
     2461 tcagcagtca caggcaaatg tctgagaaca ttagtgggac atacaggtgg agtatggtca
     2521 tcacaaatga gagacaacat catcattagt ggatctacag atcggacact caaagtgtgg
     2581 aatgcagaga ctggagaatg tatacacacc ttatatgggc atacttccac tgtgcgttgt
     2641 atgcatcttc atgaaaaaag agttgttagc ggttctcgag atgccactct tagggtttgg
     2701 gatattgaga caggccagtg tttacatgtt ttgatgggtc atgttgcagc agtccgctgt
     2761 gttcaatatg atggcaggag ggttgttagt ggagcatatg attttatggt aaaggtgtgg
     2821 gatccagaga ctgaaacctg tctacacacg ttgcaggggc atactaatag agtctattca
     2881 ttacagtttg atggtatcca tgtggtgagt ggatctcttg atacatcaat ccgtgtttgg
     2941 gatgtggaga cagggaattg cattcacacg ttaacagggc accagtcgtt aacaagtgga
     3001 atggaactca aagacaatat tcttgtctct gggaatgcag attctacagt taaaatctgg
     3061 gatatcaaaa caggacagtg tttacaaaca ttgcaaggtc ccaacaagca tcagagtgct
     3121 gtgacctgtt tacagttcaa caagaacttt gtaattacca gctcagatga tggaactgta
     3181 aaactatggg acttgaaaac gggtgaattt attcgaaacc tagtcacatt ggagagtggg
     3241 gggagtgggg gagttgtgtg gcggatcaga gcctcaaaca caaagctggt gtgtgcagtt
     3301 gggagtcgga atgggactga agaaaccaag ctgctggtgc tggactttga tgtggacatg
     3361 aagggctcga gcggtgggag ctccggtgtg agcggctggc ggctgttcaa gaagattagc
     3421 taagtttaaa cggccgcgac tctagactgc aggcatgcaa gctgatttgg ctgctgccac
     3481 cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     3541 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3601 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3661 tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt
     3721 ggtaaaatct taattaagac ttctaatata acttcgtata gcatacatta tacgaagtta
     3781 tggttccact gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     3841 ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg cacgcaggtt
     3901 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3961 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4021 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4081 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4141 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4201 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4261 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4321 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4381 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4441 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4501 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4561 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4621 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa atgaccgacc
     4681 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4741 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4801 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4861 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4921 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4981 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5041 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5101 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5161 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5221 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5281 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5341 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5401 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5461 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5521 atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt
     5581 tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag agggcagcaa
     5641 ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg ctttgatccc
     5701 aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt
     5761 ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag cagcagtgcc
     5821 cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga caccagtgaa
     5881 gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag agatggggat
     5941 gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag acaaaggctt
     6001 ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta ggcctccaaa
     6061 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     6121 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     6181 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     6241 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     6301 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     6361 acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag ttcctctttc
     6421 agaggttatt tcaggccatg gtgctgcgc
//
