LOCUS       sequence-423305-        7490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DCAF12. N-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7490)
  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 7490)
  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: pHTN-HaloTag_DCAF12:1-453
FEATURES             Location/Qualifiers
     source          1..7490
                     /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      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(3441..3459)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3491..3612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3502..3521)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3556..3575
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3857..4214
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3857..4048
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3880..3900)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4065..4200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4265..5059
                     /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(4319..4338)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4929..4948
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5123..5171
                     /note="synthetic polyadenylation signal"
     promoter        5327..5431
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5432..5500,5501..6292)
                     /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(5650..5669)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6450..7038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6530..6549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7154..7437
                     /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 gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgcttccga attcatggcc cggaaagtag ttagcaggaa
     2041 gcggaaagcg cccgcctcgc cgggagctgg gagcgacgct cagggcccgc agtttggctg
     2101 ggatcactcg cttcacaaaa ggaaaagact tcctcctgtg aagagatcct tagtatacta
     2161 cttgaagaac cgggaagtca ggctacagaa tgaaaccagc tactctcgag tgttgcatgg
     2221 ttatgcagca cagcaacttc ccagtctcct gaaggagaga gagtttcacc ttgggaccct
     2281 taataaagtg tttgcatctc agtggttgaa tcataggcaa gtggtgtgtg gcacaaaatg
     2341 caacacgcta tttgtcgtag atgtccagac aagccagatc accaagatcc ccattctgaa
     2401 agaccaggag cctggaggtg tgacccagca gggctgtggt atccatgcca tcgagctgaa
     2461 tccttctaga acactgctag ccactggagg agacaacccc aacagtcttg ccatctatcg
     2521 actacctacg ctggatcctg tgtgtgtagg agatgatgga cacaaggact ggatcttttc
     2581 catcgcatgg atcagcgaca ctatggcagt gtctggctca cgtgatggtt ctatgggact
     2641 ctgggaggtg acagatgatg ttttgaccaa aagtgatgcg agacacaatg tgtcacgggt
     2701 ccctgtgtat gcacacatca ctcacaaggc cttaaaggac atccccaaag aagacacaaa
     2761 ccctgacaac tgcaaggttc gggctctggc cttcaacaac aagaacaagg aactgggagc
     2821 agtgtctctg gatggctact ttcatctctg gaaggctgaa aatacactat ctaagctcct
     2881 ctccaccaaa ctgccatatt gccgtgagaa tgtgtgtctg gcttatggta gtgaatggtc
     2941 agtttatgca gtgggctccc aagctcatgt ctccttcttg gatccacggc agccatcata
     3001 caacgtcaag tctgtctgtt ccagggagcg aggcagtgga atccggtcag tgagtttcta
     3061 cgagcacatc atcactgtgg gaacagggca gggctccctg ctgttctatg acatccgagc
     3121 tcagagattt ctggaagaga ggctctcagc ttgttatggg tccaagccca gactagcagg
     3181 ggagaatctg aaactaacca ctggcaaagg ctggctgaat catgatgaaa cctggaggaa
     3241 ttacttttca gacattgact tcttccccaa tgctgtttac acccactgct acgactcgtc
     3301 tggaacgaaa ctctttgtgg caggaggtcc cctcccttca gggctccatg gaaactatgc
     3361 tgggctctgg agttgatgat ctagatttgg gcccaattcc tgcaggattt tgcggccgct
     3421 tgctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gccgcttcga
     3481 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3541 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3601 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     3661 tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa
     3721 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3781 tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa
     3841 cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3901 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3961 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4021 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4081 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4141 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4201 taggcttttg caaaaagctt gattcttctg acacaacagt ctcgaactta aggctagagc
     4261 caccatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4321 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4381 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4441 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4501 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4561 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4621 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4681 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4741 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4801 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4861 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4921 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4981 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5041 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5101 aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     5161 ttttttgtgt gaatcgatag cgataaggat cctctttgcg cttgcgtttt cccttgtcca
     5221 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     5281 cgtaatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5341 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5401 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5461 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5521 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5581 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5641 caaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5701 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5761 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5821 cactgcggcc aacttacttc tgacaactat cggaggaccg aaggagctaa ccgctttttt
     5881 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5941 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6001 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6061 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6121 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6181 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6241 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aattcgaaat
     6301 gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat acggttatcc
     6361 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6421 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6481 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6541 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6601 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6661 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6721 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6781 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6841 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6901 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6961 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7021 agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7081 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7141 atccttttat agtccggaaa tacaggaacg cacgctggat ggcccttcgc tgggatggtg
     7201 aaaccatgaa aaatggcagc ttcagtggat taagtggggg taatgtggcc tgtaccctct
     7261 ggttgcatag gtattcatac ggttaaaatt tatcaggcgc gattgcggca gtttttcggg
     7321 tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt
     7381 gcgtacaatt aagggattat ggtaaatcca cttactgtct gccctcgtag ccatcgagat
     7441 aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg atcgagatct
//
