LOCUS       sequence-423306-        7502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DCAF12. C-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7502)
  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 7502)
  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_DCAF12:1-453
FEATURES             Location/Qualifiers
     source          1..7502
                     /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"
     CDS             2456..2476
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2492..3376
                     /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(3448..3466)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3503..3624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3514..3533)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3568..3587
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3869..4226
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3869..4060
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3892..3912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4077..4212
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4277..5071
                     /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(4331..4350)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4941..4960
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5135..5183
                     /note="synthetic polyadenylation signal"
     promoter        5339..5443
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5444..5512,5513..6304)
                     /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(5662..5681)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6462..7050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6542..6561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7166..7449
                     /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 catggcccgg aaagtagtta gcaggaagcg gaaagcgccc gcctcgccgg gagctgggag
     1141 cgacgctcag ggcccgcagt ttggctggga tcactcgctt cacaaaagga aaagacttcc
     1201 tcctgtgaag agatccttag tatactactt gaagaaccgg gaagtcaggc tacagaatga
     1261 aaccagctac tctcgagtgt tgcatggtta tgcagcacag caacttccca gtctcctgaa
     1321 ggagagagag tttcaccttg ggacccttaa taaagtgttt gcatctcagt ggttgaatca
     1381 taggcaagtg gtgtgtggca caaaatgcaa cacgctattt gtcgtagatg tccagacaag
     1441 ccagatcacc aagatcccca ttctgaaaga ccaggagcct ggaggtgtga cccagcaggg
     1501 ctgtggtatc catgccatcg agctgaatcc ttctagaaca ctgctagcca ctggaggaga
     1561 caaccccaac agtcttgcca tctatcgact acctacgctg gatcctgtgt gtgtaggaga
     1621 tgatggacac aaggactgga tcttttccat cgcatggatc agcgacacta tggcagtgtc
     1681 tggctcacgt gatggttcta tgggactctg ggaggtgaca gatgatgttt tgaccaaaag
     1741 tgatgcgaga cacaatgtgt cacgggtccc tgtgtatgca cacatcactc acaaggcctt
     1801 aaaggacatc cccaaagaag acacaaaccc tgacaactgc aaggttcggg ctctggcctt
     1861 caacaacaag aacaaggaac tgggagcagt gtctctggat ggctactttc atctctggaa
     1921 ggctgaaaat acactatcta agctcctctc caccaaactg ccatattgcc gtgagaatgt
     1981 gtgtctggct tatggtagtg aatggtcagt ttatgcagtg ggctcccaag ctcatgtctc
     2041 cttcttggat ccacggcagc catcatacaa cgtcaagtct gtctgttcca gggagcgagg
     2101 cagtggaatc cggtcagtga gtttctacga gcacatcatc actgtgggaa cagggcaggg
     2161 ctccctgctg ttctatgaca tccgagctca gagatttctg gaagagaggc tctcagcttg
     2221 ttatgggtcc aagcccagac tagcagggga gaatctgaaa ctaaccactg gcaaaggctg
     2281 gctgaatcat gatgaaacct ggaggaatta cttttcagac attgacttct tccccaatgc
     2341 tgtttacacc cactgctacg actcgtctgg aacgaaactc tttgtggcag gaggtcccct
     2401 cccttcaggg ctccatggaa actatgctgg gctctggagt ctcgagccaa ccactgagga
     2461 tctgtacttt cagagcgata acgatggatc cgaaatcggt actggctttc cattcgaccc
     2521 ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg
     2581 cacccctgtg ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat
     2641 cccgcatgtt gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc
     2701 cgacaaacca gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat
     2761 cgaagccctg ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg
     2821 tttccactgg gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat
     2881 ccgccctatc ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt
     2941 ccgcaccacc gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac
     3001 gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc
     3061 gttcctgaat cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc
     3121 cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc
     3181 ccctgtcccg aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc
     3241 cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa
     3301 tctgctgcaa gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtctac
     3361 tctggagatt tccggttaat agaattggca tgcaagctga tccggctgct aacaaagccc
     3421 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     3481 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3541 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3601 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3661 cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc
     3721 gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     3781 ctgtgcggta tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct
     3841 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
     3901 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3961 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4021 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4081 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4141 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4201 tttttggagg cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact
     4261 taaggctaga gccaccatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4321 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4381 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4441 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4501 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4561 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4621 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4681 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4741 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4801 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4861 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4921 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4981 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5041 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5101 caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca
     5161 tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg atcctctttg cgcttgcgtt
     5221 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg
     5281 actggctttc tacgtaatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5341 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5401 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5461 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5521 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5581 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5641 gatgagcact ttcaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5701 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5761 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5821 catgagtgat aacactgcgg ccaacttact tctgacaact atcggaggac cgaaggagct
     5881 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5941 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6001 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6061 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6121 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6181 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6241 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6301 gtaattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta
     6361 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6421 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6481 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6541 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6601 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6661 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6721 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6781 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6841 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6901 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6961 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7021 cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct
     7081 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7141 atcttcacct agatcctttt atagtccgga aatacaggaa cgcacgctgg atggcccttc
     7201 gctgggatgg tgaaaccatg aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg
     7261 cctgtaccct ctggttgcat aggtattcat acggttaaaa tttatcaggc gcgattgcgg
     7321 cagtttttcg ggtggtttgt tgccattttt acctgtctgc tgccgtgatc gcgctgaacg
     7381 cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt ctgccctcgt
     7441 agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga ggatcgagat
     7501 ct
//
