LOCUS       sequence-425983-        6405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW7. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6405)
  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 6405)
  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_FBXW7:1-707
FEATURES             Location/Qualifiers
     source          1..6405
                     /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    3468..3589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3479..3498)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3704..3737)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3747..3794
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3826..4620
                     /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(3880..3899)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4490..4509
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4778..5366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4858..4877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5492..5540
                     /note="synthetic polyadenylation signal"
     CDS             complement(5564..5962)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5959..5968
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5993..6350)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6007..6142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6159..6350
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6307..6327
                     /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 tgaatcagga
     1261 actgctctct gtgggcagca aaagacgacg aactggaggc tctctgagag gtaacccttc
     1321 ctcaagccag gtagatgaag aacagatgaa tcgtgtggta gaggaggaac agcaacagca
     1381 actcagacaa caagaggagg agcacactgc aaggaatggt gaagttgttg gagtagaacc
     1441 tagacctgga ggccaaaatg attcccagca aggacagttg gaagaaaaca ataatagatt
     1501 tatttcggta gatgaggact cctcaggaaa ccaagaagaa caagaggaag atgaagaaca
     1561 tgctggtgaa caagatgagg aggatgagga ggaggaggag atggaccagg agagtgacga
     1621 ttttgatcag tctgatgata gtagcagaga agatgaacat acacatacta acagtgtcac
     1681 gaactccagt agtattgtgg acctgcccgt tcaccaactc tcctccccat tctatacaaa
     1741 aacaacaaaa atgaaaagaa agttggacca tggttctgag gtccgctctt tttctttggg
     1801 aaagaaacca tgcaaagtct cagaatatac aagtaccact gggcttgtac catgttcagc
     1861 aacaccaaca acttttgggg acctcagagc agccaatggc caagggcaac aacgacgccg
     1921 aattacatct gtccagccac ctacaggcct ccaggaatgg ctaaaaatgt ttcagagctg
     1981 gagtggacca gagaaattgc ttgctttaga tgaactcatt gatagttgtg aaccaacaca
     2041 agtaaaacat atgatgcaag tgatagaacc ccagtttcaa cgagacttca tttcattgct
     2101 ccctaaagag ttggcactct atgtgctttc attcctggaa cccaaagacc tgctacaagc
     2161 agctcagaca tgtcgctact ggagaatttt ggctgaagac aaccttctct ggagagagaa
     2221 atgcaaagaa gaggggattg atgaaccatt gcacatcaag agaagaaaag taataaaacc
     2281 aggtttcata cacagtccat ggaaaagtgc atacatcaga cagcacagaa ttgatactaa
     2341 ctggaggcga ggagaactca aatctcctaa ggtgctgaaa ggacatgatg atcatgtgat
     2401 cacatgctta cagttttgtg gtaaccgaat agttagtggt tctgatgaca acactttaaa
     2461 agtttggtca gcagtcacag gcaaatgtct gagaacatta gtgggacata caggtggagt
     2521 atggtcatca caaatgagag acaacatcat cattagtgga tctacagatc ggacactcaa
     2581 agtgtggaat gcagagactg gagaatgtat acacacctta tatgggcata cttccactgt
     2641 gcgttgtatg catcttcatg aaaaaagagt tgttagcggt tctcgagatg ccactcttag
     2701 ggtttgggat attgagacag gccagtgttt acatgttttg atgggtcatg ttgcagcagt
     2761 ccgctgtgtt caatatgatg gcaggagggt tgttagtgga gcatatgatt ttatggtaaa
     2821 ggtgtgggat ccagagactg aaacctgtct acacacgttg caggggcata ctaatagagt
     2881 ctattcatta cagtttgatg gtatccatgt ggtgagtgga tctcttgata catcaatccg
     2941 tgtttgggat gtggagacag ggaattgcat tcacacgtta acagggcacc agtcgttaac
     3001 aagtggaatg gaactcaaag acaatattct tgtctctggg aatgcagatt ctacagttaa
     3061 aatctgggat atcaaaacag gacagtgttt acaaacattg caaggtccca acaagcatca
     3121 gagtgctgtg acctgtttac agttcaacaa gaactttgta attaccagct cagatgatgg
     3181 aactgtaaaa ctatgggact tgaaaacggg tgaatttatt cgaaacctag tcacattgga
     3241 gagtgggggg agtgggggag ttgtgtggcg gatcagagcc tcaaacacaa agctggtgtg
     3301 tgcagttggg agtcggaatg ggactgaaga aaccaagctg ctggtgctgg actttgatgt
     3361 ggacatgaag taatctagag gtcaagcttc gggggtttaa acctgcaggc atgcaagctg
     3421 atttggctgc tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga
     3481 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3541 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3601 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     3661 aaacctctac aaatgtggta aaatcttaat taagacttct aatataactt cgtatagcat
     3721 acattatacg aagttatggt tccactgttg acaattaatc atcggcatag tatatcggca
     3781 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat
     3841 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3901 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3961 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4021 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4081 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4141 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4201 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4261 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4321 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4381 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4441 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4501 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4561 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4621 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     4681 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4741 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4801 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4861 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4921 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4981 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5041 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5101 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5161 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5221 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5281 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5341 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     5401 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5461 tcacctagat ccttttatag tccggaatcg tcacacaaaa aaccaacaca cagatgtaat
     5521 gaaaataaag atattttatt gcggccatcg tgatggctag agtttagccc tcccacacat
     5581 aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca
     5641 ctatggcttt gatcccagga tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc
     5701 tcaagatgcc cctgttctca tttccgatcg cgacgataca agtcaggttg ccagctgccg
     5761 cagcagcagc agtgcccagc accacgagtt ctgcacaagg tcccccagta aaatgatata
     5821 cattgacacc agtgaagatg cggccgtcgc tagagagagc tgcgctggcg acgctgtagt
     5881 cttcagagat ggggatgctg ttgattgtag ccgttgctct ttcaatgagg gtggattctt
     5941 cttgagacaa aggcttggcc atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa
     6001 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     6061 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     6121 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     6181 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     6241 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     6301 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag aaggtaccta
     6361 accaagttcc tctttcagag gttatttcag gccatggtgc tgcgc
//
