LOCUS       sequence-423291-        8079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW7. C-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8079)
  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 8079)
  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-C_FBXW7:1-707
FEATURES             Location/Qualifiers
     source          1..8079
                     /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             3254..3766
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3842..3860)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3897..4018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3908..3927)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3962..3981
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4263..4620
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4263..4454
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4286..4306)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4471..4606
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4651..5688
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5667..5675
                     /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    5712..5760
                     /note="synthetic polyadenylation signal"
     promoter        5916..6020
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6021..6089,6090..6881)
                     /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(6239..6258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7039..7627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7119..7138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7743..8026
                     /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 ggctagcaaa gcgatcgctt ccgaattcgc
     1081 caccatgaat caggaactgc tctctgtggg cagcaaaaga cgacgaactg gaggctctct
     1141 gagaggtaac ccttcctcaa gccaggtaga tgaagaacag atgaatcgtg tggtagagga
     1201 ggaacagcaa cagcaactca gacaacaaga ggaggagcac actgcaagga atggtgaagt
     1261 tgttggagta gaacctagac ctggaggcca aaatgattcc cagcaaggac agttggaaga
     1321 aaacaataat agatttattt cggtagatga ggactcctca ggaaaccaag aagaacaaga
     1381 ggaagatgaa gaacatgctg gtgaacaaga tgaggaggat gaggaggagg aggagatgga
     1441 ccaggagagt gacgattttg atcagtctga tgatagtagc agagaagatg aacatacaca
     1501 tactaacagt gtcacgaact ccagtagtat tgtggacctg cccgttcacc aactctcctc
     1561 cccattctat acaaaaacaa caaaaatgaa aagaaagttg gaccatggtt ctgaggtccg
     1621 ctctttttct ttgggaaaga aaccatgcaa agtctcagaa tatacaagta ccactgggct
     1681 tgtaccatgt tcagcaacac caacaacttt tggggacctc agagcagcca atggccaagg
     1741 gcaacaacga cgccgaatta catctgtcca gccacctaca ggcctccagg aatggctaaa
     1801 aatgtttcag agctggagtg gaccagagaa attgcttgct ttagatgaac tcattgatag
     1861 ttgtgaacca acacaagtaa aacatatgat gcaagtgata gaaccccagt ttcaacgaga
     1921 cttcatttca ttgctcccta aagagttggc actctatgtg ctttcattcc tggaacccaa
     1981 agacctgcta caagcagctc agacatgtcg ctactggaga attttggctg aagacaacct
     2041 tctctggaga gagaaatgca aagaagaggg gattgatgaa ccattgcaca tcaagagaag
     2101 aaaagtaata aaaccaggtt tcatacacag tccatggaaa agtgcataca tcagacagca
     2161 cagaattgat actaactgga ggcgaggaga actcaaatct cctaaggtgc tgaaaggaca
     2221 tgatgatcat gtgatcacat gcttacagtt ttgtggtaac cgaatagtta gtggttctga
     2281 tgacaacact ttaaaagttt ggtcagcagt cacaggcaaa tgtctgagaa cattagtggg
     2341 acatacaggt ggagtatggt catcacaaat gagagacaac atcatcatta gtggatctac
     2401 agatcggaca ctcaaagtgt ggaatgcaga gactggagaa tgtatacaca ccttatatgg
     2461 gcatacttcc actgtgcgtt gtatgcatct tcatgaaaaa agagttgtta gcggttctcg
     2521 agatgccact cttagggttt gggatattga gacaggccag tgtttacatg ttttgatggg
     2581 tcatgttgca gcagtccgct gtgttcaata tgatggcagg agggttgtta gtggagcata
     2641 tgattttatg gtaaaggtgt gggatccaga gactgaaacc tgtctacaca cgttgcaggg
     2701 gcatactaat agagtctatt cattacagtt tgatggtatc catgtggtga gtggatctct
     2761 tgatacatca atccgtgttt gggatgtgga gacagggaat tgcattcaca cgttaacagg
     2821 gcaccagtcg ttaacaagtg gaatggaact caaagacaat attcttgtct ctgggaatgc
     2881 agattctaca gttaaaatct gggatatcaa aacaggacag tgtttacaaa cattgcaagg
     2941 tcccaacaag catcagagtg ctgtgacctg tttacagttc aacaagaact ttgtaattac
     3001 cagctcagat gatggaactg taaaactatg ggacttgaaa acgggtgaat ttattcgaaa
     3061 cctagtcaca ttggagagtg gggggagtgg gggagttgtg tggcggatca gagcctcaaa
     3121 cacaaagctg gtgtgtgcag ttgggagtcg gaatgggact gaagaaacca agctgctggt
     3181 gctggacttt gatgtggaca tgaagtctag atttgggccc aattcctgca ggcgagctct
     3241 cggctcgagc ggcgtcttca cactcgaaga tttcgttggg gactggcgac agacagccgg
     3301 ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg
     3361 ggtgtccgta actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga
     3421 catccatgtc atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa
     3481 aatttttaag gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg
     3541 cacactggta atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga
     3601 aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa
     3661 caaaattatc gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat
     3721 caacggagtg accggctggc ggctgtgcga acgcattctg gcgtaaggcc gcgactctag
     3781 ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     3841 accgctgagc aataactagc ataacccctt ggggcggccg cttcgagcag acatgataag
     3901 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3961 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4021 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt
     4081 aagcaagtaa aacctctaca aatgtggtaa aatcgaattt taacaaaata ttaacgctta
     4141 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     4201 cggatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc
     4261 ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4321 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4381 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4441 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4501 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4561 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4621 aagctcgatt cttctgacac tagcgccacc atgaagaagc ccgaactcac cgctaccagc
     4681 gttgaaaaat ttctcatcga gaagttcgac agtgtgagcg acctgatgca gttgtcggag
     4741 ggcgaagaga gccgagcctt cagcttcgat gtcggcggac gcggctatgt actgcgggtg
     4801 aatagctgcg ctgatggctt ctacaaagac cgctacgtgt accgccactt cgccagcgct
     4861 gcactaccca tccccgaagt gttggacatc ggcgagttca gcgagagcct gacatactgc
     4921 atcagtagac gcgcccaagg cgttactctc caagacctcc ccgaaacaga gctgcctgct
     4981 gtgttacagc ctgtcgccga agctatggat gctattgccg ccgccgacct cagtcaaacc
     5041 agcggcttcg gcccattcgg gccccaaggc atcggccagt acacaacctg gcgggatttc
     5101 atttgcgcca ttgctgatcc ccatgtctac cactggcaga ccgtgatgga cgacaccgtg
     5161 tccgccagcg tagctcaagc cctggacgaa ctgatgctgt gggccgaaga ctgtcccgag
     5221 gtgcgccacc tcgtccatgc cgacttcggc agcaacaacg tcctgaccga caacggccgc
     5281 atcaccgccg taatcgactg gtccgaagct atgttcgggg acagtcagta cgaggtggcc
     5341 aacatcttct tctggcggcc ctggctggct tgcatggagc agcagactcg ctacttcgag
     5401 cgccggcatc ccgagctggc cggcagccct cgtctgcgag cctacatgct gcgcatcggc
     5461 ctggatcagc tctaccagag cctcgtggac ggcaacttcg acgatgctgc ctgggctcaa
     5521 ggccgctgcg atgccatcgt ccgcagcggg gccggcaccg tcggtcgcac acaaatcgct
     5581 cgccggagcg cagccgtatg gaccgacggc tgcgtcgagg tgctggccga cagcggcaac
     5641 cgccggccca gtacacgacc gcgcgctaag gaggtaggtc gagtttagac tctagccatc
     5701 acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
     5761 aatcgatagc gataaggatc ctctttgcgc ttgcgttttc ccttgtccag atagcccagt
     5821 agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac gtaatggttt
     5881 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5941 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6001 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6061 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6121 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6181 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttc aaagttctgc
     6241 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6301 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6361 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6421 acttacttct gacaactatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6481 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6541 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6601 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6661 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6721 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6781 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6841 agatcgctga gataggtgcc tcactgatta agcattggta attcgaaatg accgaccaag
     6901 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6961 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7021 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7081 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7141 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7201 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7261 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7321 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7381 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7441 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7501 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7561 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7621 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7681 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     7741 gtccggaaat acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa
     7801 aatggcagct tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg
     7861 tattcatacg gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc
     7921 catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta
     7981 agggattatg gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta
     8041 ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
