LOCUS       sequence-423421-        5687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DCAF12. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5687)
  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 5687)
  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-C_DCAF12:1-453
FEATURES             Location/Qualifiers
     source          1..5687
                     /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    2750..2871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2761..2780)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2986..3019)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3029..3076
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3108..3902
                     /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(3162..3181)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3772..3791
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4060..4648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4140..4159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4774..4822
                     /note="synthetic polyadenylation signal"
     CDS             complement(4846..5244)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5241..5250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5275..5632)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5289..5424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5441..5632
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5589..5609
                     /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 ataggggcga tcgcaacaag cttcgggaag
     1201 atctggagct agcggcgggg gatccggtgg gaattcgaca ccatggcccg gaaagtagtt
     1261 agcaggaagc ggaaagcgcc cgcctcgccg ggagctggga gcgacgctca gggcccgcag
     1321 tttggctggg atcactcgct tcacaaaagg aaaagacttc ctcctgtgaa gagatcctta
     1381 gtatactact tgaagaaccg ggaagtcagg ctacagaatg aaaccagcta ctctcgagtg
     1441 ttgcatggtt atgcagcaca gcaacttccc agtctcctga aggagagaga gtttcacctt
     1501 gggaccctta ataaagtgtt tgcatctcag tggttgaatc ataggcaagt ggtgtgtggc
     1561 acaaaatgca acacgctatt tgtcgtagat gtccagacaa gccagatcac caagatcccc
     1621 attctgaaag accaggagcc tggaggtgtg acccagcagg gctgtggtat ccatgccatc
     1681 gagctgaatc cttctagaac actgctagcc actggaggag acaaccccaa cagtcttgcc
     1741 atctatcgac tacctacgct ggatcctgtg tgtgtaggag atgatggaca caaggactgg
     1801 atcttttcca tcgcatggat cagcgacact atggcagtgt ctggctcacg tgatggttct
     1861 atgggactct gggaggtgac agatgatgtt ttgaccaaaa gtgatgcgag acacaatgtg
     1921 tcacgggtcc ctgtgtatgc acacatcact cacaaggcct taaaggacat ccccaaagaa
     1981 gacacaaacc ctgacaactg caaggttcgg gctctggcct tcaacaacaa gaacaaggaa
     2041 ctgggagcag tgtctctgga tggctacttt catctctgga aggctgaaaa tacactatct
     2101 aagctcctct ccaccaaact gccatattgc cgtgagaatg tgtgtctggc ttatggtagt
     2161 gaatggtcag tttatgcagt gggctcccaa gctcatgtct ccttcttgga tccacggcag
     2221 ccatcataca acgtcaagtc tgtctgttcc agggagcgag gcagtggaat ccggtcagtg
     2281 agtttctacg agcacatcat cactgtggga acagggcagg gctccctgct gttctatgac
     2341 atccgagctc agagatttct ggaagagagg ctctcagctt gttatgggtc caagcccaga
     2401 ctagcagggg agaatctgaa actaaccact ggcaaaggct ggctgaatca tgatgaaacc
     2461 tggaggaatt acttttcaga cattgacttc ttccccaatg ctgtttacac ccactgctac
     2521 gactcgtctg gaacgaaact ctttgtggca ggaggtcccc tcccttcagg gctccatgga
     2581 aactatgctg ggctctggag tggctcgagc ggtgggagct ccggtgtgag cggctggcgg
     2641 ctgttcaaga agattagcta agtttaaacg gccgcgactc tagactgcag gcatgcaagc
     2701 tgatttggct gctgccaccg ctgagcaagg ccgcttcgag cagacatgat aagatacatt
     2761 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2821 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2881 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag
     2941 taaaacctct acaaatgtgg taaaatctta attaagactt ctaatataac ttcgtatagc
     3001 atacattata cgaagttatg gttccactgt tgacaattaa tcatcggcat agtatatcgg
     3061 catagtataa tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag
     3121 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3181 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3241 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     3301 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3361 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3421 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3481 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3541 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3601 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     3661 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3721 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3781 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     3841 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     3901 gattcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat
     3961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4501 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4621 gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga
     4681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4741 cttcacctag atccttttat agtccggaat cgtcacacaa aaaaccaaca cacagatgta
     4801 atgaaaataa agatatttta ttgcggccat cgtgatggct agagtttagc cctcccacac
     4861 ataaccagag ggcagcaatt cacgaatccc aactgccgtc ggctgtccat cactgtcctt
     4921 cactatggct ttgatcccag gatgcagatc gagaagcacc tgtcggcacc gtccgcaggg
     4981 gctcaagatg cccctgttct catttccgat cgcgacgata caagtcaggt tgccagctgc
     5041 cgcagcagca gcagtgccca gcaccacgag ttctgcacaa ggtcccccag taaaatgata
     5101 tacattgaca ccagtgaaga tgcggccgtc gctagagaga gctgcgctgg cgacgctgta
     5161 gtcttcagag atggggatgc tgttgattgt agccgttgct ctttcaatga gggtggattc
     5221 ttcttgagac aaaggcttgg ccatggtggc gctagtgtca gaagaatcga gctttttgca
     5281 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     5341 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     5401 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     5461 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     5521 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5581 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc agaaggtacc
     5641 taaccaagtt cctctttcag aggttatttc aggccatggt gctgcgc
//
