LOCUS       sequence-425989-        7556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXO3. C-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7556)
  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 7556)
  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_FBXO3:1-471
FEATURES             Location/Qualifiers
     source          1..7556
                     /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      1080..1089
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2510..2530
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2546..3430
                     /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(3502..3520)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3557..3678
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3568..3587)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3622..3641
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3923..4280
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3923..4114
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3946..3966)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4131..4266
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4331..5125
                     /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(4385..4404)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4995..5014
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5189..5237
                     /note="synthetic polyadenylation signal"
     promoter        5393..5497
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5498..5566,5567..6358)
                     /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(5716..5735)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6516..7104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6596..6615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7220..7503
                     /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 catggcggcc atggagaccg agacggcgcc gctgacccta gagtcgctgc ccaccgatcc
     1141 cctgctcctc atcttatcct ttttggacta tcgggatcta atcaactgtt gttatgtcag
     1201 tcgaagactt agccagctat caagtcatga tccgctgtgg agaagacatt gcaaaaaata
     1261 ctggctgata tctgaggaag agaaaacaca gaagaatcag tgttggaaat ctctcttcat
     1321 agatacttac tctgatgtag gaagatacat tgaccattat gctgctatta aaaaggcctg
     1381 ggatgatctc aagaaatatt tggagcccag gtgtcctcgg atggttttat ctctgaaaga
     1441 gggtgctcga gaggaagacc tcgatgctgt ggaagcgcag attggctgca agcttcctga
     1501 cgattatcga tgttcatacc gaattcacaa tggacagaag ttagtggttc ctgggttatt
     1561 gggaagcatg gcactgtcta atcactatcg ttctgaagat ttgttagacg tcgatacagc
     1621 tgccggagga ttccagcaga gacagggact gaaatactgt ctccctttaa ctttttgcat
     1681 acatactggt ttgagtcagt acatagcagt ggaagctgca gagggccgaa acaaaaatga
     1741 agttttctac caatgtccag accaaatggc tcgaaatcca gctgctattg acatgtttat
     1801 tataggtgct acttttactg actggtttac ctcttatgtc aaaaatgttg tatcaggtgg
     1861 cttccccatc atcagagacc aaattttcag atatgttcac gatccagaat gtgtagcaac
     1921 aactggggat attactgtgt cagtttccac atcgtttctg ccagaactta gctctgtaca
     1981 tccaccccac tatttcttca cataccgaat caggattgaa atgtcaaaag atgcacttcc
     2041 tgagaaggcc tgtcagttgg acagtcgcta ttggagaata acaaatgcta agggtgacgt
     2101 ggaagaagtt caaggacctg gagtagttgg tgaatttcca atcatcagcc caggtcgggt
     2161 atatgaatac acaagctgta ccacattctc tacaacatca ggatacatgg aaggatatta
     2221 taccttccat tttctttact ttaaagacaa gatctttaat gttgccattc cccgattcca
     2281 tatggcatgt ccaacattca gggtgtctat agcccgattg gaaatgggtc ctgatgaata
     2341 tgaagagatg gaagaagagg aggaggagga agaggaggaa gacgaggatg atgattcagc
     2401 agatatggat gaatcagatg aagatgatga agaggagaga cggaggagag tctttgatgt
     2461 tcccattcgc agacgccgct gctcacgcct ttttctcgag ccaaccactg aggatctgta
     2521 ctttcagagc gataacgatg gatccgaaat cggtactggc tttccattcg acccccatta
     2581 tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg atggcacccc
     2641 tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca tcatcccgca
     2701 tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca aatccgacaa
     2761 accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct tcatcgaagc
     2821 cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc tgggtttcca
     2881 ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt tcatccgccc
     2941 tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg ccttccgcac
     3001 caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg gtacgctgcc
     3061 gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg agccgttcct
     3121 gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa tcgccggtga
     3181 gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc agtcccctgt
     3241 cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg aagccgctcg
     3301 cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc tgaatctgct
     3361 gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt ctactctgga
     3421 gatttccggt taatagaatt ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg
     3481 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcggccg
     3541 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3601 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3661 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3721 gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcgaattt
     3781 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     3841 ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata acctctgaaa
     3901 gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3961 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4021 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4081 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4141 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4201 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4261 gaggcctagg cttttgcaaa aagcttgatt cttctgacac aacagtctcg aacttaaggc
     4321 tagagccacc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4381 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4441 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4501 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4561 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4621 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4681 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4741 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4801 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4861 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4921 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4981 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5041 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5101 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5161 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     5221 tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct
     5281 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc
     5341 tttctacgta atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5401 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5461 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5521 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5581 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5641 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5701 cactttcaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5761 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5821 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5881 tgataacact gcggccaact tacttctgac aactatcgga ggaccgaagg agctaaccgc
     5941 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6001 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6061 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6121 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6181 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6241 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6301 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaatt
     6361 cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg
     6421 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6481 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6541 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6601 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6661 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6721 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6781 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6841 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6901 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6961 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7021 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7081 acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct
     7141 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7201 acctagatcc ttttatagtc cggaaataca ggaacgcacg ctggatggcc cttcgctggg
     7261 atggtgaaac catgaaaaat ggcagcttca gtggattaag tgggggtaat gtggcctgta
     7321 ccctctggtt gcataggtat tcatacggtt aaaatttatc aggcgcgatt gcggcagttt
     7381 ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt
     7441 agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc tcgtagccat
     7501 cgagataaac cgcagtactc cggccacgat gcgtccggcg tagaggatcg agatct
//
