LOCUS       sequence-426256-        7829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW5. N-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7829)
  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 7829)
  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: pHTN-HaloTag_FBXW5:1-566
FEATURES             Location/Qualifiers
     source          1..7829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..60
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        41..59
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      69..78
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             75..965
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             978..998
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(2788..2806)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2838..2959
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2849..2868)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2903..2922
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3204..3561
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3204..3395
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3227..3247)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3412..3547
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3612..4406
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3666..3685)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4276..4295
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4470..4518
                     /note="synthetic polyadenylation signal"
     promoter        4674..4778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4779..4847,4848..5639)
                     /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(4997..5016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5797..6385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5877..5896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6501..6784
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        6975..7354
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7355..7558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7509..7529
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          7694..7826
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     7809..7828
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 tccactccca gttcaattac agctcttaag gctagagtat taatacgact cactataggg
       61 ctagcaaagc caccatggca gaaatcggta ctggctttcc attcgacccc cattatgtgg
      121 aagtcctggg cgagcgcatg cactacgtcg atgttggtcc gcgcgatggc acccctgtgc
      181 tgttcctgca cggtaacccg acctcctcct acgtgtggcg caacatcatc ccgcatgttg
      241 caccgaccca tcgctgcatt gctccagacc tgatcggtat gggcaaatcc gacaaaccag
      301 acctgggtta tttcttcgac gaccacgtcc gcttcatgga tgccttcatc gaagccctgg
      361 gtctggaaga ggtcgtcctg gtcattcacg actggggctc cgctctgggt ttccactggg
      421 ccaagcgcaa tccagagcgc gtcaaaggta ttgcatttat ggagttcatc cgccctatcc
      481 cgacctggga cgaatggcca gaatttgccc gcgagacctt ccaggccttc cgcaccaccg
      541 acgtcggccg caagctgatc atcgatcaga acgtttttat cgagggtacg ctgccgatgg
      601 gtgtcgtccg cccgctgact gaagtcgaga tggaccatta ccgcgagccg ttcctgaatc
      661 ctgttgaccg cgagccactg tggcgcttcc caaacgagct gccaatcgcc ggtgagccag
      721 cgaacatcgt cgcgctggtc gaagaataca tggactggct gcaccagtcc cctgtcccga
      781 agctgctgtt ctggggcacc ccaggcgttc tgatcccacc ggccgaagcc gctcgcctgg
      841 ccaaaagcct gcctaactgc aaggctgtgg acatcggccc gggtctgaat ctgctgcaag
      901 aagacaaccc ggacctgatc ggcagcgaga tcgcgcgctg gctgtcgacg ctcgagattt
      961 ccggcgagcc aaccactgag gatctgtact ttcagagcga taacgcgatc gcttccgaat
     1021 tcatggacga gggcggcacg cccctgctcc ccgacagcct ggtctaccag atcttcctga
     1081 gcctgggccc ggccgacgtg ctggccgccg ggctggtgtg ccgccaatgg caggccgtgt
     1141 cgcgggacga gttcctgtgg agggagcagt tctaccgcta ctaccaggtg gcccgcgacg
     1201 tgccccgaca cccagcggcc atgtcctggt acgaggagtt ccagcggctg tatgacacgg
     1261 tgccctgcgt ggaggtgcag acgctgcggg aacacacaga ccaggtcctg cacctcagct
     1321 tctcccattc cgggtaccag ttcgcgtcct gctccaagga ctgcactgtg aagatctgga
     1381 gcaacgacct gaccatctcg ctgctgcaca gcgcggacat gcggccctac aactggagct
     1441 acacccagtt ctcccagttc aacaaggacg actcgctact gctggcctcg ggggtgttcc
     1501 tggggccgca caactcctca tccggcgaga ttgctgtcat cagcctagac tccttcgcgc
     1561 tgctgtcccg cgtgcggaac aagccctatg acgtgtttgg ctgttggctc accgagacca
     1621 gcctcatctc ggggaacctg caccgcatcg gagatatcac ctcctgctcg gtgctgtggc
     1681 tcaacaatgc cttccaggat gtggagtcag agaacgtcaa cgtggtgaag cggctgttca
     1741 agatccagaa cctcaatgcc agcaccgtcc gcacggtgat ggtggccgac tgcagccgct
     1801 tcgacagccc tgacctgctg ctggaagccg gtgacccggc cacgtccccc tgccgcatct
     1861 ttgacctggg cagcgacaac gaggaggtgg tggctggccc ggcccccgcc cacgccaagg
     1921 agggcttgcg gcactttctg gaccgcgtgc tggaggggcg ggcgcagcca cagctgtcgg
     1981 agcgcatgct agagaccaag gtggccgagc tgctggccca gggccacacc aagccacccg
     2041 agcgcagtgc cacaggcgcc aagagcaagt acctcatctt caccactggc tgcctcacct
     2101 actccccaca ccagatcggc atcaagcaga tcctgccaca ccagatgacc acggcagggc
     2161 ccgtgctggg tgagggccgg ggctccgatg ccttcttcga cgcgctggac cacgtcatag
     2221 acatacacgg acacatcatc ggcatgggcc tgtcgcccga caacaggtac ctgtacgtga
     2281 acagccgcgc ctggcccaac ggtgcggtgg tggccgaccc catgcagccg ccaccaatcg
     2341 cggaggagat tgacctgctg gtgttcgacc tcaagaccat gcgggaggtg aggcgggctc
     2401 tgcgtgcaca ccgcgcctac acgcccaacg acgagtgctt cttcatcttc ctggacgtca
     2461 gcagggactt cgtggccagc ggggcggagg accggcacgg ctacatctgg gaccgccact
     2521 acaacatctg tctggccagg ctgcggcacg aggatgtggt caactcagtg gtcttcagtc
     2581 cccaggagca ggagctgctg ctcacggcca gcgacgacgc caccatcaaa gcctggcgct
     2641 ccccacgcac catgcgcgtc ctccaggcac ctcgcccacg gcctcgcacc ttcttctcct
     2701 ggcttgccag ccagaggcgc tgatgatcta gatttgggcc caattcctgc aggattttgc
     2761 ggccgcttgc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttggcc
     2821 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2881 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2941 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3001 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcgaatt
     3061 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3121 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa
     3181 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3241 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3301 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3361 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3421 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3481 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3541 ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaacttaagg
     3601 ctagagccac catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3661 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3721 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3781 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3841 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3901 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3961 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4021 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4081 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4141 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4201 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4261 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4321 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4381 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4441 gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt
     4501 gtgttggttt tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt gcgttttccc
     4561 ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg
     4621 ctttctacgt aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4681 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4741 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4801 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4861 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4921 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4981 gcactttcaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5041 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5101 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5161 gtgataacac tgcggccaac ttacttctga caactatcgg aggaccgaag gagctaaccg
     5221 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5281 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5341 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5401 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5461 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5521 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5581 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaat
     5641 tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg
     5701 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5761 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5821 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5881 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5941 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6001 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6061 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6121 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6181 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6241 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6301 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6361 tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6421 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6481 cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc ccttcgctgg
     6541 gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa tgtggcctgt
     6601 accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat tgcggcagtt
     6661 tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct gaacgcgttt
     6721 tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca
     6781 tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatcttca
     6841 atattggcca ttagccatat tattcattgg ttatatagca taaatcaata ttggctattg
     6901 gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat
     6961 atgaccgcca tgttggcatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     7021 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     7081 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     7141 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     7201 cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg
     7261 taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca
     7321 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa
     7381 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     7441 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc
     7501 cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     7561 tttagtgaac cgtcagatca ctagaagctt tattgcggta gtttatcaca gttaaattgc
     7621 taacgcagtc agtgcttctg acacaacagt ctcgaactta agctgcagaa gttggtcgtg
     7681 aggcactggg caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact
     7741 gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga
     7801 catccacttt gcctttctct ccacaggtg
//
