LOCUS       sequence-423286-        8028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW7. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8028)
  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 8028)
  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: pNLF1-N_FBXW7:1-707
FEATURES             Location/Qualifiers
     source          1..8028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3796..3814)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3846..3967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3857..3876)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3911..3930
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4212..4569
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4212..4403
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4235..4255)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4420..4555
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4600..5637
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5616..5624
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5661..5709
                     /note="synthetic polyadenylation signal"
     promoter        5865..5969
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5970..6038,6039..6830)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6188..6207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6988..7576)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7068..7087)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7692..7975
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgcttc cgaattcatg aatcaggaac
     1621 tgctctctgt gggcagcaaa agacgacgaa ctggaggctc tctgagaggt aacccttcct
     1681 caagccaggt agatgaagaa cagatgaatc gtgtggtaga ggaggaacag caacagcaac
     1741 tcagacaaca agaggaggag cacactgcaa ggaatggtga agttgttgga gtagaaccta
     1801 gacctggagg ccaaaatgat tcccagcaag gacagttgga agaaaacaat aatagattta
     1861 tttcggtaga tgaggactcc tcaggaaacc aagaagaaca agaggaagat gaagaacatg
     1921 ctggtgaaca agatgaggag gatgaggagg aggaggagat ggaccaggag agtgacgatt
     1981 ttgatcagtc tgatgatagt agcagagaag atgaacatac acatactaac agtgtcacga
     2041 actccagtag tattgtggac ctgcccgttc accaactctc ctccccattc tatacaaaaa
     2101 caacaaaaat gaaaagaaag ttggaccatg gttctgaggt ccgctctttt tctttgggaa
     2161 agaaaccatg caaagtctca gaatatacaa gtaccactgg gcttgtacca tgttcagcaa
     2221 caccaacaac ttttggggac ctcagagcag ccaatggcca agggcaacaa cgacgccgaa
     2281 ttacatctgt ccagccacct acaggcctcc aggaatggct aaaaatgttt cagagctgga
     2341 gtggaccaga gaaattgctt gctttagatg aactcattga tagttgtgaa ccaacacaag
     2401 taaaacatat gatgcaagtg atagaacccc agtttcaacg agacttcatt tcattgctcc
     2461 ctaaagagtt ggcactctat gtgctttcat tcctggaacc caaagacctg ctacaagcag
     2521 ctcagacatg tcgctactgg agaattttgg ctgaagacaa ccttctctgg agagagaaat
     2581 gcaaagaaga ggggattgat gaaccattgc acatcaagag aagaaaagta ataaaaccag
     2641 gtttcataca cagtccatgg aaaagtgcat acatcagaca gcacagaatt gatactaact
     2701 ggaggcgagg agaactcaaa tctcctaagg tgctgaaagg acatgatgat catgtgatca
     2761 catgcttaca gttttgtggt aaccgaatag ttagtggttc tgatgacaac actttaaaag
     2821 tttggtcagc agtcacaggc aaatgtctga gaacattagt gggacataca ggtggagtat
     2881 ggtcatcaca aatgagagac aacatcatca ttagtggatc tacagatcgg acactcaaag
     2941 tgtggaatgc agagactgga gaatgtatac acaccttata tgggcatact tccactgtgc
     3001 gttgtatgca tcttcatgaa aaaagagttg ttagcggttc tcgagatgcc actcttaggg
     3061 tttgggatat tgagacaggc cagtgtttac atgttttgat gggtcatgtt gcagcagtcc
     3121 gctgtgttca atatgatggc aggagggttg ttagtggagc atatgatttt atggtaaagg
     3181 tgtgggatcc agagactgaa acctgtctac acacgttgca ggggcatact aatagagtct
     3241 attcattaca gtttgatggt atccatgtgg tgagtggatc tcttgataca tcaatccgtg
     3301 tttgggatgt ggagacaggg aattgcattc acacgttaac agggcaccag tcgttaacaa
     3361 gtggaatgga actcaaagac aatattcttg tctctgggaa tgcagattct acagttaaaa
     3421 tctgggatat caaaacagga cagtgtttac aaacattgca aggtcccaac aagcatcaga
     3481 gtgctgtgac ctgtttacag ttcaacaaga actttgtaat taccagctca gatgatggaa
     3541 ctgtaaaact atgggacttg aaaacgggtg aatttattcg aaacctagtc acattggaga
     3601 gtggggggag tgggggagtt gtgtggcgga tcagagcctc aaacacaaag ctggtgtgtg
     3661 cagttgggag tcggaatggg actgaagaaa ccaagctgct ggtgctggac tttgatgtgg
     3721 acatgaagta ataatctaga tttgggccca attcctgcag gattttgcgg ccgcttgctg
     3781 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggccgc ttcgagcaga
     3841 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3901 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3961 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     4021 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat
     4081 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4141 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     4201 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4261 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4321 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4381 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4441 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4501 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4561 ttttgcaaaa agctcgattc ttctgacact agcgccacca tgaagaagcc cgaactcacc
     4621 gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga cctgatgcag
     4681 ttgtcggagg gcgaagagag ccgagccttc agcttcgatg tcggcggacg cggctatgta
     4741 ctgcgggtga atagctgcgc tgatggcttc tacaaagacc gctacgtgta ccgccacttc
     4801 gccagcgctg cactacccat ccccgaagtg ttggacatcg gcgagttcag cgagagcctg
     4861 acatactgca tcagtagacg cgcccaaggc gttactctcc aagacctccc cgaaacagag
     4921 ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg ctattgccgc cgccgacctc
     4981 agtcaaacca gcggcttcgg cccattcggg ccccaaggca tcggccagta cacaacctgg
     5041 cgggatttca tttgcgccat tgctgatccc catgtctacc actggcagac cgtgatggac
     5101 gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg ggccgaagac
     5161 tgtcccgagg tgcgccacct cgtccatgcc gacttcggca gcaacaacgt cctgaccgac
     5221 aacggccgca tcaccgccgt aatcgactgg tccgaagcta tgttcgggga cagtcagtac
     5281 gaggtggcca acatcttctt ctggcggccc tggctggctt gcatggagca gcagactcgc
     5341 tacttcgagc gccggcatcc cgagctggcc ggcagccctc gtctgcgagc ctacatgctg
     5401 cgcatcggcc tggatcagct ctaccagagc ctcgtggacg gcaacttcga cgatgctgcc
     5461 tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt cggtcgcaca
     5521 caaatcgctc gccggagcgc agccgtatgg accgacggct gcgtcgaggt gctggccgac
     5581 agcggcaacc gccggcccag tacacgaccg cgcgctaagg aggtaggtcg agtttagact
     5641 ctagccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     5701 ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga
     5761 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
     5821 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5881 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5941 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6001 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6061 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6121 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca
     6181 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6241 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6301 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6361 ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc
     6421 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6481 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6541 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6601 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6661 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6721 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6781 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga
     6841 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6901 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6961 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7021 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7081 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7141 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7201 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7261 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7321 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7381 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7441 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7501 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7561 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7621 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7681 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     7741 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     7801 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     7861 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     7921 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     7981 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
