LOCUS       sequence-425904-        5466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human FBXW12 isoform 2. 
            N-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5466)
  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 5466)
  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: pBiT3.1-N_FBXW12:1-394
FEATURES             Location/Qualifiers
     source          1..5466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2529..2650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2540..2559)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2765..2798)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2808..2855
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2887..3681
                     /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(2941..2960)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3551..3570
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3839..4427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3919..3938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4553..4601
                     /note="synthetic polyadenylation signal"
     CDS             complement(4625..5023)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5020..5029
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5054..5411)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5068..5203
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5220..5411
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5368..5388
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggta tggagatccg
     1261 attgcctgac ttagctttga agcgaatctt ctctttcctg gacctgttcg gcttgctgca
     1321 ggtttcccag gtgaacaagc attggaatag gattgcagac agtgattacc tgtggaggtc
     1381 actatctctg cagagatggg actgcagcaa cttcaccaat caacacctgg gcacacacac
     1441 atggaagcaa tttttcctgc atcaaagaag gaaggagctt cggttggcat tggcacagcc
     1501 gcataacttt atctacaaag taactaagaa catcgcattt gagacggagt tggcttatct
     1561 ctcaggaaat agacttacag tggatgaaca ggagaaatca atcatttgta gtgtatctcc
     1621 aaagcaagag ctttgtgcct gggatgtgca agaggttggc gatgctgcag gtgacatcta
     1681 cacatttaca ctgcctgggt taagagatgt ttctaaagtt actgcatttc aatatggtat
     1741 tgtacttcta cactgctctc ctgacaagaa atgggtattt gcatgtggga catacagtcg
     1801 taccttgcca caggtattcc tcacagagtc cttactgaga ccatcagaag gcagtgttcc
     1861 tctgtctacc tttctcccac ataaattatg tgccagcgcc tgctggaccc caaaggtgaa
     1921 aaacaggata acactgatgt cccaaagtag cactggaaaa aagacagaat ttatcacctt
     1981 tgatctaaca accaagaaga ctggaggcca aacagtcatc caagcatatg agatcgcaag
     2041 tttccaggtg gcagctcatc tgaagtgccc tatctggatg ggagccagtg atggatatat
     2101 gattgtcttt accagtggac catacttgtt actcttcagc atcactggct tcctgctgca
     2161 acgatttgag gaccatcagg cagccatcaa caacttctgg gtggatcctt gctatgtgct
     2221 caccacatcc gagaactctg tgcacgtgta catgtgggaa gaaggaggcc gccatccata
     2281 cctcaggagc tgctgtcacc tggaaaacac gtggcatgat cacacaacag acagctgcat
     2341 ctccagtgtg atgtgtgata atgcaagcat agtacttagg gtgaggaaag taagtgactc
     2401 cagcattctg gtgatgtatt ctttgaatac gtaatctaga ggtcaagctt cgggggttta
     2461 aacctgcagg catgcaagct gatttggctg ctgccaccgc tgagcaaggc cgcttcgagc
     2521 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2581 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2641 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     2701 ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcttaa ttaagacttc
     2761 taatataact tcgtatagca tacattatac gaagttatgg ttccactgtt gacaattaat
     2821 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa acccaggagg
     2881 cagatcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2941 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3001 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3061 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3121 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3181 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3241 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3301 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3361 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     3421 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3481 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3541 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3601 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3661 cttcttgacg agttcttctg attcgaaatg accgaccaag cgacgcccaa ccggtatcag
     3721 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3781 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3841 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3901 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3961 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4021 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4081 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4141 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4201 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4261 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4321 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4381 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa gatcctttga
     4441 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4501 tgagattatc aaaaaggatc ttcacctaga tccttttata gtccggaatc gtcacacaaa
     4561 aaaccaacac acagatgtaa tgaaaataaa gatattttat tgcggccatc gtgatggcta
     4621 gagtttagcc ctcccacaca taaccagagg gcagcaattc acgaatccca actgccgtcg
     4681 gctgtccatc actgtccttc actatggctt tgatcccagg atgcagatcg agaagcacct
     4741 gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc atttccgatc gcgacgatac
     4801 aagtcaggtt gccagctgcc gcagcagcag cagtgcccag caccacgagt tctgcacaag
     4861 gtcccccagt aaaatgatat acattgacac cagtgaagat gcggccgtcg ctagagagag
     4921 ctgcgctggc gacgctgtag tcttcagaga tggggatgct gttgattgta gccgttgctc
     4981 tttcaatgag ggtggattct tcttgagaca aaggcttggc catggtggcg ctagtgtcag
     5041 aagaatcgag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     5101 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat
     5161 ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg
     5221 ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     5281 ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc
     5341 ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac agctggttct
     5401 ttccgcctca gaaggtacct aaccaagttc ctctttcaga ggttatttca ggccatggtg
     5461 ctgcgc
//
