LOCUS       sequence-423348-        8402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human SUZ12. C-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8402)
  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 8402)
  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_SUZ12:1-739
FEATURES             Location/Qualifiers
     source          1..8402
                     /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      1079..1088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3356..3376
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             3392..4276
                     /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(4348..4366)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4403..4524
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4414..4433)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4468..4487
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4769..5126
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4769..4960
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4792..4812)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4977..5112
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5177..5971
                     /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(5231..5250)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5841..5860
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6035..6083
                     /note="synthetic polyadenylation signal"
     promoter        6239..6343
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6344..6412,6413..7204)
                     /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(6562..6581)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7362..7950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7442..7461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8066..8349
                     /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 ccgaattcgc
     1081 caccatggcg cctcagaagc acggcggtgg gggagggggc ggctcggggc ccagcgcggg
     1141 gtccggggga ggcggcttcg ggggttcggc ggcggtggcg gcggcgacgg cttcgggcgg
     1201 caaatccggc ggcgggagct gtggaggggg tggcagttac tcggcctcct cctcctcctc
     1261 cgcggcggca gcggcggggg ctgcggtgtt accggtgaag aagccgaaaa tggagcacgt
     1321 ccaggctgac cacgagcttt tcctccaggc ctttgagaag ccaacacaga tctatagatt
     1381 tcttcgaact cggaatctca tagcaccaat atttttgcac agaactctta cttacatgtc
     1441 tcatcgaaac tccagaacaa acatcaaaag gaaaacattt aaagttgatg atatgttatc
     1501 aaaagtagag aaaatgaaag gagagcaaga atctcatagc ttgtcagctc atttgcagct
     1561 tacgtttact ggtttcttcc acaaaaatga taagccatca ccaaactcag aaaatgaaca
     1621 aaattctgtt accctggaag tcctgcttgt gaaagtttgc cacaaaaaaa gaaaggatgt
     1681 aagttgtcca ataaggcaag ttcccacagg taaaaagcag gtgcctttga atcctgacct
     1741 caatcaaaca aaacccggaa atttcccgtc ccttgcagtt tccagtaatg aatttgaacc
     1801 tagtaacagc catatggtga agtcttactc gttgctattt agagtgactc gtccaggaag
     1861 aagagagttt aatggaatga ttaatggaga aaccaatgaa aatattgatg tcaatgaaga
     1921 gcttccagcc agaagaaaac gaaatcgtga ggatggggaa aagacatttg ttgcacaaat
     1981 gacagtattt gataaaaaca ggcgcttaca gcttttagat ggggaatatg aagtagccat
     2041 gcaggaaatg gaagaatgtc caataagcaa gaaaagagca acatgggaga ctattcttga
     2101 tgggaagagg ctgcctccat tcgaaacatt ttctcaggga cctacgttgc agttcactct
     2161 tcgttggaca ggagagacca atgataaatc tacggctcct attgccaaac ctcttgccac
     2221 tagaaattca gagagtctcc atcaggaaaa caagcctggt tcagttaaac ctactcaaac
     2281 tattgctgtt aaagaatcat tgactacaga tctacaaaca agaaaagaaa aggatactcc
     2341 aaatgaaaac cgacaaaaat taagaatatt ttatcagttt ctctataaca acaatacaag
     2401 gcaacaaact gaagcaagag atgacctgca ttgcccttgg tgtactctga actgccgcaa
     2461 actttatagt ttactcaagc atcttaaact ctgccatagc agatttatct tcaactatgt
     2521 ttatcatcca aaaggtgcta ggatagatgt ttctatcaat gagtgttatg atggctccta
     2581 tgcaggaaat cctcaggata ttcatcgcca acctggattt gcttttagtc gcaacggacc
     2641 agttaagaga acacctatca cacatattct tgtgtgcagg ccaaaacgaa caaaagcaag
     2701 catgtctgaa tttcttgaat ctgaagatgg ggaagtagaa cagcaaagaa catatagtag
     2761 tggccacaat cgtctgtatt tccatagtga tacctgctta cctctccgtc cacaagaaat
     2821 ggaagtagat agtgaagatg aaaaggatcc tgaatggcta agagaaaaaa ccattacaca
     2881 aattgaagag ttttctgatg ttaatgaagg agagaaagaa gtgatgaaac tctggaatct
     2941 ccatgtcatg aagcatgggt ttattgctga caatcaaatg aatcatgcct gtatgctgtt
     3001 tgtagaaaat tatggacaga aaataattaa gaagaattta tgtcgaaact tcatgcttca
     3061 tctagtcagc atgcatgact ttaatcttat tagcataatg tcaatagata aagctgttac
     3121 caagctccgt gaaatgcagc aaaaattaga aaagggggaa tctgcttccc ctgcaaacga
     3181 agaaataact gaagaacaaa atgggacagc aaatggattt agtgaaatta actcaaaaga
     3241 gaaagctttg gaaacagata gtgtctcagg ggtttcaaaa cagagcaaaa aacaaaaact
     3301 ccggatatct agatttgggc ccaattcctg caggcgagct ctcgagccaa ccactgagga
     3361 tctgtacttt cagagcgata acgatggatc cgaaatcggt actggctttc cattcgaccc
     3421 ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg
     3481 cacccctgtg ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat
     3541 cccgcatgtt gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc
     3601 cgacaaacca gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat
     3661 cgaagccctg ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg
     3721 tttccactgg gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat
     3781 ccgccctatc ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt
     3841 ccgcaccacc gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac
     3901 gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc
     3961 gttcctgaat cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc
     4021 cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc
     4081 ccctgtcccg aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc
     4141 cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa
     4201 tctgctgcaa gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtctac
     4261 tctggagatt tccggttaat agaattggca tgcaagctga tccggctgct aacaaagccc
     4321 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     4381 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     4441 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     4501 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     4561 cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc
     4621 gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     4681 ctgtgcggta tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct
     4741 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
     4801 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4861 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4921 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4981 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5041 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5101 tttttggagg cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact
     5161 taaggctaga gccaccatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5221 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5281 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5341 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5401 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5461 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5521 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5581 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5641 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5701 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5761 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5821 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5881 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5941 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6001 caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca
     6061 tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg atcctctttg cgcttgcgtt
     6121 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg
     6181 actggctttc tacgtaatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     6241 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6301 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6361 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6421 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6481 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6541 gatgagcact ttcaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6601 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6661 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6721 catgagtgat aacactgcgg ccaacttact tctgacaact atcggaggac cgaaggagct
     6781 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6841 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6901 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6961 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7021 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7081 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7141 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7201 gtaattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta
     7261 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7321 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7381 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7441 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7501 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7561 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7621 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7681 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7741 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7801 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7861 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7921 cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct
     7981 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     8041 atcttcacct agatcctttt atagtccgga aatacaggaa cgcacgctgg atggcccttc
     8101 gctgggatgg tgaaaccatg aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg
     8161 cctgtaccct ctggttgcat aggtattcat acggttaaaa tttatcaggc gcgattgcgg
     8221 cagtttttcg ggtggtttgt tgccattttt acctgtctgc tgccgtgatc gcgctgaacg
     8281 cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt ctgccctcgt
     8341 agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga ggatcgagat
     8401 ct
//
