LOCUS       sequence-423320-        7841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW5. C-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7841)
  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 7841)
  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: pHTC-HaloTag_FBXW5:1-566
FEATURES             Location/Qualifiers
     source          1..7841
                     /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             2795..2815
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2831..3715
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(3787..3805)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3842..3963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3853..3872)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3907..3926
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4208..4565
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4208..4399
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4231..4251)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4416..4551
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4616..5410
                     /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(4670..4689)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5280..5299
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5474..5522
                     /note="synthetic polyadenylation signal"
     promoter        5678..5782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5783..5851,5852..6643)
                     /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(6001..6020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6801..7389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6881..6900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7505..7788
                     /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 ccgaattcac
     1081 catggacgag ggcggcacgc ccctgctccc cgacagcctg gtctaccaga tcttcctgag
     1141 cctgggcccg gccgacgtgc tggccgccgg gctggtgtgc cgccaatggc aggccgtgtc
     1201 gcgggacgag ttcctgtgga gggagcagtt ctaccgctac taccaggtgg cccgcgacgt
     1261 gccccgacac ccagcggcca tgtcctggta cgaggagttc cagcggctgt atgacacggt
     1321 gccctgcgtg gaggtgcaga cgctgcggga acacacagac caggtcctgc acctcagctt
     1381 ctcccattcc gggtaccagt tcgcgtcctg ctccaaggac tgcactgtga agatctggag
     1441 caacgacctg accatctcgc tgctgcacag cgcggacatg cggccctaca actggagcta
     1501 cacccagttc tcccagttca acaaggacga ctcgctactg ctggcctcgg gggtgttcct
     1561 ggggccgcac aactcctcat ccggcgagat tgctgtcatc agcctagact ccttcgcgct
     1621 gctgtcccgc gtgcggaaca agccctatga cgtgtttggc tgttggctca ccgagaccag
     1681 cctcatctcg gggaacctgc accgcatcgg agatatcacc tcctgctcgg tgctgtggct
     1741 caacaatgcc ttccaggatg tggagtcaga gaacgtcaac gtggtgaagc ggctgttcaa
     1801 gatccagaac ctcaatgcca gcaccgtccg cacggtgatg gtggccgact gcagccgctt
     1861 cgacagccct gacctgctgc tggaagccgg tgacccggcc acgtccccct gccgcatctt
     1921 tgacctgggc agcgacaacg aggaggtggt ggctggcccg gcccccgccc acgccaagga
     1981 gggcttgcgg cactttctgg accgcgtgct ggaggggcgg gcgcagccac agctgtcgga
     2041 gcgcatgcta gagaccaagg tggccgagct gctggcccag ggccacacca agccacccga
     2101 gcgcagtgcc acaggcgcca agagcaagta cctcatcttc accactggct gcctcaccta
     2161 ctccccacac cagatcggca tcaagcagat cctgccacac cagatgacca cggcagggcc
     2221 cgtgctgggt gagggccggg gctccgatgc cttcttcgac gcgctggacc acgtcataga
     2281 catacacgga cacatcatcg gcatgggcct gtcgcccgac aacaggtacc tgtacgtgaa
     2341 cagccgcgcc tggcccaacg gtgcggtggt ggccgacccc atgcagccgc caccaatcgc
     2401 ggaggagatt gacctgctgg tgttcgacct caagaccatg cgggaggtga ggcgggctct
     2461 gcgtgcgcac cgcgcctaca cgcccaacga cgagtgcttc ttcatcttcc tggacgtcag
     2521 cagggacttc gtggccagcg gggcggagga ccggcacggc tacatctggg accgccacta
     2581 caacatctgt ctggccaggc tgcggcacga ggatgtggtc aactcagtgg tcttcagtcc
     2641 ccaggagcag gagctgctgc tcacggccag cgacgacgcc accatcaaag cctggcgctc
     2701 cccacgcacc atgcgcgtcc tccaggcacc tcgcccacgg cctcgcacct tcttctcctg
     2761 gcttgccagc cagaggcgcc tcgagccaac cactgaggat ctgtactttc agagcgataa
     2821 cgatggatcc gaaatcggta ctggctttcc attcgacccc cattatgtgg aagtcctggg
     2881 cgagcgcatg cactacgtcg atgttggtcc gcgcgatggc acccctgtgc tgttcctgca
     2941 cggtaacccg acctcctcct acgtgtggcg caacatcatc ccgcatgttg caccgaccca
     3001 tcgctgcatt gctccagacc tgatcggtat gggcaaatcc gacaaaccag acctgggtta
     3061 tttcttcgac gaccacgtcc gcttcatgga tgccttcatc gaagccctgg gtctggaaga
     3121 ggtcgtcctg gtcattcacg actggggctc cgctctgggt ttccactggg ccaagcgcaa
     3181 tccagagcgc gtcaaaggta ttgcatttat ggagttcatc cgccctatcc cgacctggga
     3241 cgaatggcca gaatttgccc gcgagacctt ccaggccttc cgcaccaccg acgtcggccg
     3301 caagctgatc atcgatcaga acgtttttat cgagggtacg ctgccgatgg gtgtcgtccg
     3361 cccgctgact gaagtcgaga tggaccatta ccgcgagccg ttcctgaatc ctgttgaccg
     3421 cgagccactg tggcgcttcc caaacgagct gccaatcgcc ggtgagccag cgaacatcgt
     3481 cgcgctggtc gaagaataca tggactggct gcaccagtcc cctgtcccga agctgctgtt
     3541 ctggggcacc ccaggcgttc tgatcccacc ggccgaagcc gctcgcctgg ccaaaagcct
     3601 gcctaactgc aaggctgtgg acatcggccc gggtctgaat ctgctgcaag aagacaaccc
     3661 ggacctgatc ggcagcgaga tcgcgcgctg gctgtctact ctggagattt ccggttaata
     3721 gaattggcat gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     3781 ctgccaccgc tgagcaataa ctagcataac cccttggggc ggccgcttcg agcagacatg
     3841 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3901 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3961 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     4021 tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca aaatattaac
     4081 gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4141 atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     4201 gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4261 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4321 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4381 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4441 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4501 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4561 gcaaaaagct tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat
     4621 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4681 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4741 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4801 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4861 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4921 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4981 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5041 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5101 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     5161 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5221 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5281 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5341 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5401 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5461 tcacgatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     5521 tgaatcgata gcgataagga tcctctttgc gcttgcgttt tcccttgtcc agatagccca
     5581 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtaatggt
     5641 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5701 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5761 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5821 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5881 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5941 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt tcaaagttct
     6001 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6061 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6121 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6181 caacttactt ctgacaacta tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6241 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6301 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6361 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6421 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6481 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6541 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6601 acagatcgct gagataggtg cctcactgat taagcattgg taattcgaaa tgaccgacca
     6661 agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6721 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6781 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6841 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6901 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6961 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7021 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7081 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7141 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7201 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     7261 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7321 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7381 ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7441 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7501 tagtccggaa atacaggaac gcacgctgga tggcccttcg ctgggatggt gaaaccatga
     7561 aaaatggcag cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata
     7621 ggtattcata cggttaaaat ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt
     7681 gccattttta cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat
     7741 taagggatta tggtaaatcc acttactgtc tgccctcgta gccatcgaga taaaccgcag
     7801 tactccggcc acgatgcgtc cggcgtagag gatcgagatc t
//
