LOCUS       sequence-423287-        7533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW11. N-terminal NanoLuc
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7533)
  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 7533)
  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_FBXW11:1-542
FEATURES             Location/Qualifiers
     source          1..7533
                     /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(3301..3319)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3351..3472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3362..3381)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3416..3435
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3717..4074
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3717..3908
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3740..3760)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3925..4060
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4105..5142
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5121..5129
                     /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    5166..5214
                     /note="synthetic polyadenylation signal"
     promoter        5370..5474
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5475..5543,5544..6335)
                     /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(5693..5712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6493..7081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6573..6592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7197..7480
                     /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 gagcccgact
     1621 cggtgattga ggacaagacc atcgagctca tgtgttctgt gccaaggtct ttgtggctag
     1681 gctgcgccaa cctggtagag agcatgtgcg cactgagttg cctgcagagc atgcccagtg
     1741 tcagatgtct ccagataagt aatggaacat catctgtgat cgtctccaga aagaggccat
     1801 cagaaggaaa ctatcaaaaa gaaaaagact tgtgtattaa atattttgac cagtggtctg
     1861 aatcagatca agtggaattt gtggaacatc ttatttcacg aatgtgtcat tatcagcatg
     1921 gacatattaa ctcttacctg aagcccatgt tgcagcggga ctttattacc gctttaccag
     1981 agcaaggctt agatcacata gcagaaaaca ttctttcgta cctggatgcc aggtctctgt
     2041 gtgcagcaga gctggtatgt aaagaatggc agcgagtgat ctcagaagga atgctttgga
     2101 agaagctgat tgaacgaatg gtacgcactg atcccctatg gaaaggactt tcagaaagaa
     2161 gagggtggga tcagtacctg tttaaaaaca gacccacaga tggccctcca aattcatttt
     2221 ataggtcatt atacccaaag attatccagg atatagagac tatagaatct aactggcggt
     2281 gtggacgaca caacttgcag aggattcagt gccgctctga aaatagtaaa ggtgtctact
     2341 gtttacagta cgatgatgaa aaaattatca gtggcctacg agataattct attaagatat
     2401 gggataaaac cagcctggaa tgtttgaaag tgttaacagg acacacaggc tctgtcctct
     2461 gtctgcagta tgatgagcgt gtcattgtaa ctggctcttc agattctacg gtgagagtgt
     2521 gggatgtgaa cacgggtgaa gttcttaaca cattgatcca ccacaatgag gctgtattgc
     2581 acttacgctt cagcaatgga ctgatggtga cctgttccaa ggaccgctcc attgctgtgt
     2641 gggacatggc ttctgcgacc gacatcactt tacgccgtgt cctggttggc caccgggctg
     2701 ccgtcaatgt agtagacttt gacgacaagt acatcgtgtc tgcctctggt gacaggacca
     2761 tcaaagtctg gagcacgagc acctgtgaat ttgttcgtac tctcaatggg cacaagcggg
     2821 gcattgcctg tctccagtac agggatcgcc tggttgttag tggatcatca gataatacca
     2881 ttaggctctg ggatattgaa tgtggtgcct gtttaagagt cctagaggga catgaagaat
     2941 tggtccgatg catccggttt gataacaaga ggattgtcag tggggcctat gatgggaaaa
     3001 ttaaagtttg ggacttgcaa gctgctcttg accctcgagc cccagcaagc acattgtgtt
     3061 tgcgcacatt ggtggaacat tctggacgtg tgtttcggct ccagtttgat gagtttcaga
     3121 tcatcagcag ctcccatgat gacactattt tgatttggga tttcttaaat gtgcctccca
     3181 gtgcccagaa tgagacccgt tctccctcca gaacatacac ttacatctct agataataat
     3241 ctagatttgg gcccaattcc tgcaggattt tgcggccgct tgctgagttg gctgctgcca
     3301 ccgctgagca ataactagca taaccccttg gccgcttcga gcagacatga taagatacat
     3361 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3421 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3481 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa
     3541 gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg cttacaattt
     3601 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgcggatc
     3661 tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg taccttctga
     3721 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3781 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3841 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3901 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3961 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4021 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4081 gattcttctg acactagcgc caccatgaag aagcccgaac tcaccgctac cagcgttgaa
     4141 aaatttctca tcgagaagtt cgacagtgtg agcgacctga tgcagttgtc ggagggcgaa
     4201 gagagccgag ccttcagctt cgatgtcggc ggacgcggct atgtactgcg ggtgaatagc
     4261 tgcgctgatg gcttctacaa agaccgctac gtgtaccgcc acttcgccag cgctgcacta
     4321 cccatccccg aagtgttgga catcggcgag ttcagcgaga gcctgacata ctgcatcagt
     4381 agacgcgccc aaggcgttac tctccaagac ctccccgaaa cagagctgcc tgctgtgtta
     4441 cagcctgtcg ccgaagctat ggatgctatt gccgccgccg acctcagtca aaccagcggc
     4501 ttcggcccat tcgggcccca aggcatcggc cagtacacaa cctggcggga tttcatttgc
     4561 gccattgctg atccccatgt ctaccactgg cagaccgtga tggacgacac cgtgtccgcc
     4621 agcgtagctc aagccctgga cgaactgatg ctgtgggccg aagactgtcc cgaggtgcgc
     4681 cacctcgtcc atgccgactt cggcagcaac aacgtcctga ccgacaacgg ccgcatcacc
     4741 gccgtaatcg actggtccga agctatgttc ggggacagtc agtacgaggt ggccaacatc
     4801 ttcttctggc ggccctggct ggcttgcatg gagcagcaga ctcgctactt cgagcgccgg
     4861 catcccgagc tggccggcag ccctcgtctg cgagcctaca tgctgcgcat cggcctggat
     4921 cagctctacc agagcctcgt ggacggcaac ttcgacgatg ctgcctgggc tcaaggccgc
     4981 tgcgatgcca tcgtccgcag cggggccggc accgtcggtc gcacacaaat cgctcgccgg
     5041 agcgcagccg tatggaccga cggctgcgtc gaggtgctgg ccgacagcgg caaccgccgg
     5101 cccagtacac gaccgcgcgc taaggaggta ggtcgagttt agactctagc catcacgatg
     5161 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     5221 tagcgataag gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga
     5281 cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtaatg gtttcttaga
     5341 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5401 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5461 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5521 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5581 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5641 agagttttcg ccccgaagaa cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg
     5701 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5761 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5821 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5881 ttctgacaac tatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5941 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6001 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6061 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6121 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6181 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6241 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6301 ctgagatagg tgcctcactg attaagcatt ggtaattcga aatgaccgac caagcgacgc
     6361 ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6421 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6481 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6541 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6601 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6661 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6721 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6781 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6841 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6901 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6961 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7021 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca
     7081 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7141 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg
     7201 aaatacagga acgcacgctg gatggccctt cgctgggatg gtgaaaccat gaaaaatggc
     7261 agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc tctggttgca taggtattca
     7321 tacggttaaa atttatcagg cgcgattgcg gcagtttttc gggtggtttg ttgccatttt
     7381 tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca attaagggat
     7441 tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg
     7501 ccacgatgcg tccggcgtag aggatcgaga tct
//
