LOCUS       sequence-425955-        7176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXL2. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7176)
  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 7176)
  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_FBXL2:1-423
FEATURES             Location/Qualifiers
     source          1..7176
                     /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(2944..2962)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2994..3115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3005..3024)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3059..3078
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3360..3717
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3360..3551
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3383..3403)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3568..3703
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3748..4785
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4764..4772
                     /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    4809..4857
                     /note="synthetic polyadenylation signal"
     promoter        5013..5117
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5118..5186,5187..5978)
                     /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(5336..5355)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6136..6724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6216..6235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6840..7123
                     /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 gttttctcaa
     1621 acaatgatga aggccttatt aacaaaaagt tacccaaaga acttctgtta agaatatttt
     1681 ccttcttgga tatagtaact ttgtgccgat gtgcacagat ttccaaggct tggaacatct
     1741 tagccctgga tggaagcaac tggcaaagaa tagatctttt taactttcaa acagatgtag
     1801 agggtcgagt ggtggaaaat atctcgaagc gatgcggtgg attcctgagg aagctcagct
     1861 tgcgaggctg cattggtgtt ggggattcct ccttgaagac ctttgcacag aactgccgaa
     1921 acattgaaca tttgaacctc aatggatgca caaaaatcac tgacagcacg tgttatagcc
     1981 ttagcagatt ctgttccaag ctgaaacatc tggatctgac ctcctgtgtg tctattacaa
     2041 acagctcctt gaaggggatc agtgagggct gccgaaacct ggagtacctg aacctctctt
     2101 ggtgtgatca gatcacgaag gatggcatcg aggcactggt gcgaggttgt cgaggcctga
     2161 aagccctgct cctgaggggc tgcacacagt tagaagatga agctctgaaa cacattcaga
     2221 attactgcca tgagcttgtg agcctcaact tgcagtcctg ctcacgtatc acggatgaag
     2281 gtgtggtgca gatatgcagg ggctgtcacc ggctacaggc tctctgcctt tcgggttgca
     2341 gcaacctcac agatgcctct cttacagccc tgggtttgaa ctgtccgcga ctgcaaattt
     2401 tggaggctgc ccgatgctcc catttgactg acgcaggttt tacactttta gctcggaatt
     2461 gccacgaatt ggagaagatg gatcttgaag aatgcatcct gataaccgac agcacactca
     2521 tccagctctc cattcactgt cctaaactgc aagccctgag cctgtcccac tgtgaactca
     2581 tcacagatga tgggatcctg cacctgagca acagtacctg tggccatgag aggctgcggg
     2641 tactggagtt ggacaactgc ctcctcatca ctgatgtggc cctggaacac ctagagaact
     2701 gccgaggcct ggagcgcctc gagctgtacg actgccagca ggttacccgt gcaggcatca
     2761 agcggatgcg ggctcagctc cctcatgtca aagtccacgc ctactttgct cccgtcaccc
     2821 caccgacagc agtggcagga agtggacagc gactgtgcag gtgctgtgtc attctctaat
     2881 aatctagatt tgggcccaat tcctgcagga ttttgcggcc gcttgctgag ttggctgctg
     2941 ccaccgctga gcaataacta gcataacccc ttggccgctt cgagcagaca tgataagata
     3001 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3061 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3121 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg tttttttaag
     3181 caagtaaaac ctctacaaat gtggtaaaat cgaattttaa caaaatatta acgcttacaa
     3241 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgcgg
     3301 atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc
     3361 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3421 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3481 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3541 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3601 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3661 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3721 ctcgattctt ctgacactag cgccaccatg aagaagcccg aactcaccgc taccagcgtt
     3781 gaaaaatttc tcatcgagaa gttcgacagt gtgagcgacc tgatgcagtt gtcggagggc
     3841 gaagagagcc gagccttcag cttcgatgtc ggcggacgcg gctatgtact gcgggtgaat
     3901 agctgcgctg atggcttcta caaagaccgc tacgtgtacc gccacttcgc cagcgctgca
     3961 ctacccatcc ccgaagtgtt ggacatcggc gagttcagcg agagcctgac atactgcatc
     4021 agtagacgcg cccaaggcgt tactctccaa gacctccccg aaacagagct gcctgctgtg
     4081 ttacagcctg tcgccgaagc tatggatgct attgccgccg ccgacctcag tcaaaccagc
     4141 ggcttcggcc cattcgggcc ccaaggcatc ggccagtaca caacctggcg ggatttcatt
     4201 tgcgccattg ctgatcccca tgtctaccac tggcagaccg tgatggacga caccgtgtcc
     4261 gccagcgtag ctcaagccct ggacgaactg atgctgtggg ccgaagactg tcccgaggtg
     4321 cgccacctcg tccatgccga cttcggcagc aacaacgtcc tgaccgacaa cggccgcatc
     4381 accgccgtaa tcgactggtc cgaagctatg ttcggggaca gtcagtacga ggtggccaac
     4441 atcttcttct ggcggccctg gctggcttgc atggagcagc agactcgcta cttcgagcgc
     4501 cggcatcccg agctggccgg cagccctcgt ctgcgagcct acatgctgcg catcggcctg
     4561 gatcagctct accagagcct cgtggacggc aacttcgacg atgctgcctg ggctcaaggc
     4621 cgctgcgatg ccatcgtccg cagcggggcc ggcaccgtcg gtcgcacaca aatcgctcgc
     4681 cggagcgcag ccgtatggac cgacggctgc gtcgaggtgc tggccgacag cggcaaccgc
     4741 cggcccagta cacgaccgcg cgctaaggag gtaggtcgag tttagactct agccatcacg
     4801 atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
     4861 cgatagcgat aaggatcctc tttgcgcttg cgttttccct tgtccagata gcccagtagc
     4921 tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgta atggtttctt
     4981 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5041 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5101 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5161 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5221 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5281 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cactttcaaa gttctgctat
     5341 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5401 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5461 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5521 tacttctgac aactatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5581 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5641 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5701 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5761 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5821 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5881 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5941 tcgctgagat aggtgcctca ctgattaagc attggtaatt cgaaatgacc gaccaagcga
     6001 cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6061 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6121 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6181 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6241 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6301 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6361 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6421 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6481 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6541 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6601 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6661 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt
     6721 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6781 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc
     6841 cggaaataca ggaacgcacg ctggatggcc cttcgctggg atggtgaaac catgaaaaat
     6901 ggcagcttca gtggattaag tgggggtaat gtggcctgta ccctctggtt gcataggtat
     6961 tcatacggtt aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat
     7021 ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg
     7081 gattatggta aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc
     7141 cggccacgat gcgtccggcg tagaggatcg agatct
//
