LOCUS       sequence-423347-        8349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human SUZ12. N-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8349)
  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 8349)
  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_SUZ12:1-739
FEATURES             Location/Qualifiers
     source          1..8349
                     /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(4300..4318)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4350..4471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4361..4380)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4415..4434
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4716..5073
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4716..4907
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4739..4759)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4924..5059
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5124..5918
                     /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(5178..5197)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5788..5807
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5982..6030
                     /note="synthetic polyadenylation signal"
     promoter        6186..6290
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6291..6359,6360..7151)
                     /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(6509..6528)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7309..7897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7389..7408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8013..8296
                     /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 attcatggcg cctcagaagc acggcggtgg
     2041 gggagggggc ggctcggggc ccagcgcggg gtccggggga ggcggcttcg ggggttcggc
     2101 ggcggtggcg gcggcgacgg cttcgggcgg caaatccggc ggcgggagct gtggaggggg
     2161 tggcagttac tcggcctcct cctcctcctc cgcggcggca gcggcggggg ctgcggtgtt
     2221 accggtgaag aagccgaaaa tggagcacgt ccaggctgac cacgagcttt tcctccaggc
     2281 ctttgagaag ccaacacaga tctatagatt tcttcgaact cggaatctca tagcaccaat
     2341 atttttgcac agaactctta cttacatgtc tcatcgaaac tccagaacaa acatcaaaag
     2401 gaaaacattt aaagttgatg atatgttatc aaaagtagag aaaatgaaag gagagcaaga
     2461 atctcatagc ttgtcagctc atttgcagct tacgtttact ggtttcttcc acaaaaatga
     2521 taagccatca ccaaactcag aaaatgaaca aaattctgtt accctggaag tcctgcttgt
     2581 gaaagtttgc cacaaaaaaa gaaaggatgt aagttgtcca ataaggcaag ttcccacagg
     2641 taaaaagcag gtgcctttga atcctgacct caatcaaaca aaacccggaa atttcccgtc
     2701 ccttgcagtt tccagtaatg aatttgaacc tagtaacagc catatggtga agtcttactc
     2761 gttgctattt agagtgactc gtccaggaag aagagagttt aatggaatga ttaatggaga
     2821 aaccaatgaa aatattgatg tcaatgaaga gcttccagcc agaagaaaac gaaatcgtga
     2881 ggatggggaa aagacatttg ttgcacaaat gacagtattt gataaaaaca ggcgcttaca
     2941 gcttttagat ggggaatatg aagtagccat gcaggaaatg gaagaatgtc caataagcaa
     3001 gaaaagagca acatgggaga ctattcttga tgggaagagg ctgcctccat tcgaaacatt
     3061 ttctcaggga cctacgttgc agttcactct tcgttggaca ggagagacca atgataaatc
     3121 tacggctcct attgccaaac ctcttgccac tagaaattca gagagtctcc atcaggaaaa
     3181 caagcctggt tcagttaaac ctactcaaac tattgctgtt aaagaatcat tgactacaga
     3241 tctacaaaca agaaaagaaa aggatactcc aaatgaaaac cgacaaaaat taagaatatt
     3301 ttatcagttt ctctataaca acaatacaag gcaacaaact gaagcaagag atgacctgca
     3361 ttgcccttgg tgtactctga actgccgcaa actttatagt ttactcaagc atcttaaact
     3421 ctgccatagc agatttatct tcaactatgt ttatcatcca aaaggtgcta ggatagatgt
     3481 ttctatcaat gagtgttatg atggctccta tgcaggaaat cctcaggata ttcatcgcca
     3541 acctggattt gcttttagtc gcaacggacc agttaagaga acacctatca cacatattct
     3601 tgtgtgcagg ccaaaacgaa caaaagcaag catgtctgaa tttcttgaat ctgaagatgg
     3661 ggaagtagaa cagcaaagaa catatagtag tggccacaat cgtctgtatt tccatagtga
     3721 tacctgctta cctctccgtc cacaagaaat ggaagtagat agtgaagatg aaaaggatcc
     3781 tgaatggcta agagaaaaaa ccattacaca aattgaagag ttttctgatg ttaatgaagg
     3841 agagaaagaa gtgatgaaac tctggaatct ccatgtcatg aagcatgggt ttattgctga
     3901 caatcaaatg aatcatgcct gtatgctgtt tgtagaaaat tatggacaga aaataattaa
     3961 gaagaattta tgtcgaaact tcatgcttca tctagtcagc atgcatgact ttaatcttat
     4021 tagcataatg tcaatagata aagctgttac caagctccgt gaaatgcagc aaaaattaga
     4081 aaagggggaa tctgcttccc ctgcaaacga agaaataact gaagaacaaa atgggacagc
     4141 aaatggattt agtgaaatta actcaaaaga gaaagctttg gaaacagata gtgtctcagg
     4201 ggtttcaaaa cagagcaaaa aacaaaaact ctgagatatc tagatttggg cccaattcct
     4261 gcaggatttt gcggccgctt gctgagttgg ctgctgccac cgctgagcaa taactagcat
     4321 aaccccttgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     4381 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4441 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     4501 tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg
     4561 taaaatcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     4621 tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa
     4681 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
     4741 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4801 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4861 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4921 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4981 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5041 ggaggctttt ttggaggcct aggcttttgc aaaaagcttg attcttctga cacaacagtc
     5101 tcgaacttaa ggctagagcc accatgattg aacaagatgg attgcacgca ggttctccgg
     5161 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5221 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5281 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5341 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5401 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5461 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5521 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5581 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5641 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5701 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5761 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5821 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5881 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5941 gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt
     6001 cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc ctctttgcgc
     6061 ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt
     6121 tctgcggact ggctttctac gtaatggttt cttagacgtc aggtggcact tttcggggaa
     6181 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6241 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6301 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6361 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6421 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6481 ttccaatgat gagcactttc aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6541 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6601 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6661 ccataaccat gagtgataac actgcggcca acttacttct gacaactatc ggaggaccga
     6721 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6781 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6841 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6901 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6961 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     7021 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7081 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7141 agcattggta attcgaaatg accgaccaag cgacgcccaa ccggtatcag ctcactcaaa
     7201 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7741 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7861 caagcagcag attacgcgca gaaaaaaagg atttcaagaa gatcctttga tcttttctac
     7921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7981 aaaaaggatc ttcacctaga tccttttata gtccggaaat acaggaacgc acgctggatg
     8041 gcccttcgct gggatggtga aaccatgaaa aatggcagct tcagtggatt aagtgggggt
     8101 aatgtggcct gtaccctctg gttgcatagg tattcatacg gttaaaattt atcaggcgcg
     8161 attgcggcag tttttcgggt ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg
     8221 ctgaacgcgt tttagcggtg cgtacaatta agggattatg gtaaatccac ttactgtctg
     8281 ccctcgtagc catcgagata aaccgcagta ctccggccac gatgcgtccg gcgtagagga
     8341 tcgagatct
//
