LOCUS       sequence-423292-        7584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW11. C-terminal NanoLuc
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7584)
  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 7584)
  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_FBXW11:1-542
FEATURES             Location/Qualifiers
     source          1..7584
                     /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"
     regulatory      1079..1088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2759..3271
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3347..3365)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3402..3523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3413..3432)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3467..3486
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3768..4125
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3768..3959
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3791..3811)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3976..4111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4156..5193
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5172..5180
                     /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    5217..5265
                     /note="synthetic polyadenylation signal"
     promoter        5421..5525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5526..5594,5595..6386)
                     /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(5744..5763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6544..7132)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6624..6643)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7248..7531
                     /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 caccatggag cccgactcgg tgattgagga caagaccatc gagctcatgt gttctgtgcc
     1141 aaggtctttg tggctaggct gcgccaacct ggtagagagc atgtgcgcac tgagttgcct
     1201 gcagagcatg cccagtgtca gatgtctcca gataagtaat ggaacatcat ctgtgatcgt
     1261 ctccagaaag aggccatcag aaggaaacta tcaaaaagaa aaagacttgt gtattaaata
     1321 ttttgaccag tggtctgaat cagatcaagt ggaatttgtg gaacatctta tttcacgaat
     1381 gtgtcattat cagcatggac atattaactc ttacctgaag cccatgttgc agcgggactt
     1441 tattaccgct ttaccagagc aaggcttaga tcacatagca gaaaacattc tttcgtacct
     1501 ggatgccagg tctctgtgtg cagcagagct ggtatgtaaa gaatggcagc gagtgatctc
     1561 agaaggaatg ctttggaaga agctgattga acgaatggta cgcactgatc ccctatggaa
     1621 aggactttca gaaagaagag ggtgggatca gtacctgttt aaaaacagac ccacagatgg
     1681 ccctccaaat tcattttata ggtcattata cccaaagatt atccaggata tagagactat
     1741 agaatctaac tggcggtgtg gacgacacaa cttgcagagg attcagtgcc gctctgaaaa
     1801 tagtaaaggt gtctactgtt tacagtacga tgatgaaaaa attatcagtg gcctacgaga
     1861 taattctatt aagatatggg ataaaaccag cctggaatgt ttgaaagtgt taacaggaca
     1921 cacaggctct gtcctctgtc tgcagtatga tgagcgtgtc attgtaactg gctcttcaga
     1981 ttctacggtg agagtgtggg atgtgaacac gggtgaagtt cttaacacat tgatccacca
     2041 caatgaggct gtattgcact tacgcttcag caatggactg atggtgacct gttccaagga
     2101 ccgctccatt gctgtgtggg acatggcttc tgcgaccgac atcactttac gccgtgtcct
     2161 ggttggccac cgggctgccg tcaatgtagt agactttgac gacaagtaca tcgtgtctgc
     2221 ctctggtgac aggaccatca aagtctggag cacgagcacc tgtgaatttg ttcgtactct
     2281 caatgggcac aagcggggca ttgcctgtct ccagtacagg gatcgcctgg ttgttagtgg
     2341 atcatcagat aataccatta ggctctggga tattgaatgt ggtgcctgtt taagagtcct
     2401 agagggacat gaagaattgg tccgatgcat ccggtttgat aacaagagga ttgtcagtgg
     2461 ggcctatgat gggaaaatta aagtttggga cttgcaagct gctcttgacc ctcgagcccc
     2521 agcaagcaca ttgtgtttgc gcacattggt ggaacattct ggacgtgtgt ttcggctcca
     2581 gtttgatgag tttcagatca tcagcagctc ccatgatgac actattttga tttgggattt
     2641 cttaaatgtg cctcccagtg cccagaatga gacccgttct ccctccagaa catacactta
     2701 catctctaga tctagatttg ggcccaattc ctgcaggcga gctctcggct cgagcggcgt
     2761 cttcacactc gaagatttcg ttggggactg gcgacagaca gccggctaca acctggacca
     2821 agtccttgaa cagggaggtg tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc
     2881 gatccaaagg attgtcctga gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat
     2941 cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt
     3001 gtaccctgtg gatgatcatc actttaaggt gatcctgcac tatggcacac tggtaatcga
     3061 cggggttacg ccgaacatga tcgactattt cggacggccg tatgaaggca tcgccgtgtt
     3121 cgacggcaaa aagatcactg taacagggac cctgtggaac ggcaacaaaa ttatcgacga
     3181 gcgcctgatc aaccccgacg gctccctgct gttccgagta accatcaacg gagtgaccgg
     3241 ctggcggctg tgcgaacgca ttctggcgta aggccgcgac tctagggcat gcaagctgat
     3301 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     3361 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3421 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3481 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3541 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     3601 ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     3661 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca
     3721 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     3781 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3841 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3901 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3961 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4021 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4081 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cgattcttct
     4141 gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta ccagcgttga aaaatttctc
     4201 atcgagaagt tcgacagtgt gagcgacctg atgcagttgt cggagggcga agagagccga
     4261 gccttcagct tcgatgtcgg cggacgcggc tatgtactgc gggtgaatag ctgcgctgat
     4321 ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca gcgctgcact acccatcccc
     4381 gaagtgttgg acatcggcga gttcagcgag agcctgacat actgcatcag tagacgcgcc
     4441 caaggcgtta ctctccaaga cctccccgaa acagagctgc ctgctgtgtt acagcctgtc
     4501 gccgaagcta tggatgctat tgccgccgcc gacctcagtc aaaccagcgg cttcggccca
     4561 ttcgggcccc aaggcatcgg ccagtacaca acctggcggg atttcatttg cgccattgct
     4621 gatccccatg tctaccactg gcagaccgtg atggacgaca ccgtgtccgc cagcgtagct
     4681 caagccctgg acgaactgat gctgtgggcc gaagactgtc ccgaggtgcg ccacctcgtc
     4741 catgccgact tcggcagcaa caacgtcctg accgacaacg gccgcatcac cgccgtaatc
     4801 gactggtccg aagctatgtt cggggacagt cagtacgagg tggccaacat cttcttctgg
     4861 cggccctggc tggcttgcat ggagcagcag actcgctact tcgagcgccg gcatcccgag
     4921 ctggccggca gccctcgtct gcgagcctac atgctgcgca tcggcctgga tcagctctac
     4981 cagagcctcg tggacggcaa cttcgacgat gctgcctggg ctcaaggccg ctgcgatgcc
     5041 atcgtccgca gcggggccgg caccgtcggt cgcacacaaa tcgctcgccg gagcgcagcc
     5101 gtatggaccg acggctgcgt cgaggtgctg gccgacagcg gcaaccgccg gcccagtaca
     5161 cgaccgcgcg ctaaggaggt aggtcgagtt tagactctag ccatcacgat ggccgcaata
     5221 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa
     5281 ggatcctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     5341 ggggtcagca ccgtttctgc ggactggctt tctacgtaat ggtttcttag acgtcaggtg
     5401 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5461 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5521 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5581 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5641 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5701 gccccgaaga acgttttcca atgatgagca ctttcaaagt tctgctatgt ggcgcggtat
     5761 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5821 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5881 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5941 ctatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6001 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6061 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6121 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6181 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6241 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6301 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6361 gtgcctcact gattaagcat tggtaattcg aaatgaccga ccaagcgacg cccaaccggt
     6421 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6481 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6541 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6601 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6661 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6721 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6781 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6841 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6901 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6961 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7021 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7081 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     7141 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7201 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg
     7261 aacgcacgct ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt
     7321 ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa
     7381 aatttatcag gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct
     7441 gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa
     7501 tccacttact gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc
     7561 gtccggcgta gaggatcgag atct
//
