LOCUS       sequence-425897-        5643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DCAF12 N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5643)
  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 5643)
  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_DCAF12:1-453
FEATURES             Location/Qualifiers
     source          1..5643
                     /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    2706..2827
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2717..2736)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2942..2975)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2985..3032
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3064..3858
                     /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(3118..3137)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3728..3747
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4016..4604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4096..4115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4730..4778
                     /note="synthetic polyadenylation signal"
     CDS             complement(4802..5200)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5197..5206
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5231..5588)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5245..5380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5397..5588
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5545..5565
                     /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 tggcccggaa
     1261 agtagttagc aggaagcgga aagcgcccgc ctcgccggga gctgggagcg acgctcaggg
     1321 cccgcagttt ggctgggatc actcgcttca caaaaggaaa agacttcctc ctgtgaagag
     1381 atccttagta tactacttga agaaccggga agtcaggcta cagaatgaaa ccagctactc
     1441 tcgagtgttg catggttatg cagcacagca acttcccagt ctcctgaagg agagagagtt
     1501 tcaccttggg acccttaata aagtgtttgc atctcagtgg ttgaatcata ggcaagtggt
     1561 gtgtggcaca aaatgcaaca cgctatttgt cgtagatgtc cagacaagcc agatcaccaa
     1621 gatccccatt ctgaaagacc aggagcctgg aggtgtgacc cagcagggct gtggtatcca
     1681 tgccatcgag ctgaatcctt ctagaacact gctagccact ggaggagaca accccaacag
     1741 tcttgccatc tatcgactac ctacgctgga tcctgtgtgt gtaggagatg atggacacaa
     1801 ggactggatc ttttccatcg catggatcag cgacactatg gcagtgtctg gctcacgtga
     1861 tggttctatg ggactctggg aggtgacaga tgatgttttg accaaaagtg atgcgagaca
     1921 caatgtgtca cgggtccctg tgtatgcaca catcactcac aaggccttaa aggacatccc
     1981 caaagaagac acaaaccctg acaactgcaa ggttcgggct ctggccttca acaacaagaa
     2041 caaggaactg ggagcagtgt ctctggatgg ctactttcat ctctggaagg ctgaaaatac
     2101 actatctaag ctcctctcca ccaaactgcc atattgccgt gagaatgtgt gtctggctta
     2161 tggtagtgaa tggtcagttt atgcagtggg ctcccaagct catgtctcct tcttggatcc
     2221 acggcagcca tcatacaacg tcaagtctgt ctgttccagg gagcgaggca gtggaatccg
     2281 gtcagtgagt ttctacgagc acatcatcac tgtgggaaca gggcagggct ccctgctgtt
     2341 ctatgacatc cgagctcaga gatttctgga agagaggctc tcagcttgtt atgggtccaa
     2401 gcccagacta gcaggggaga atctgaaact aaccactggc aaaggctggc tgaatcatga
     2461 tgaaacctgg aggaattact tttcagacat tgacttcttc cccaatgctg tttacaccca
     2521 ctgctacgac tcgtctggaa cgaaactctt tgtggcagga ggtcccctcc cttcagggct
     2581 ccatggaaac tatgctgggc tctggagtta atctagaggt caagcttcgg gggtttaaac
     2641 ctgcaggcat gcaagctgat ttggctgctg ccaccgctga gcaaggccgc ttcgagcaga
     2701 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2761 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2821 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     2881 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcttaatta agacttctaa
     2941 tataacttcg tatagcatac attatacgaa gttatggttc cactgttgac aattaatcat
     3001 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag
     3061 atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3121 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3181 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3241 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3301 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3361 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3421 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3481 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3541 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3601 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3661 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3721 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3781 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3841 cttgacgagt tcttctgatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc
     3901 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3961 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4021 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4081 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4141 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4201 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4261 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4321 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4381 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4441 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4501 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4561 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     4621 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4681 gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaatcgtc acacaaaaaa
     4741 ccaacacaca gatgtaatga aaataaagat attttattgc ggccatcgtg atggctagag
     4801 tttagccctc ccacacataa ccagagggca gcaattcacg aatcccaact gccgtcggct
     4861 gtccatcact gtccttcact atggctttga tcccaggatg cagatcgaga agcacctgtc
     4921 ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt tccgatcgcg acgatacaag
     4981 tcaggttgcc agctgccgca gcagcagcag tgcccagcac cacgagttct gcacaaggtc
     5041 ccccagtaaa atgatataca ttgacaccag tgaagatgcg gccgtcgcta gagagagctg
     5101 cgctggcgac gctgtagtct tcagagatgg ggatgctgtt gattgtagcc gttgctcttt
     5161 caatgagggt ggattcttct tgagacaaag gcttggccat ggtggcgcta gtgtcagaag
     5221 aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     5281 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     5341 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     5401 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     5461 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     5521 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc tggttctttc
     5581 cgcctcagaa ggtacctaac caagttcctc tttcagaggt tatttcaggc catggtgctg
     5641 cgc
//
