LOCUS       sequence-191925-        8805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human EZH2 with HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8805)
  AUTHORS   Bracken AP, Pasini D, Capra M, Prosperini E, Colli E, Helin K
  TITLE     EZH2 is downstream of the pRB-E2F pathway, essential for 
            proliferation and amplified in cancer.
  JOURNAL   EMBO J. 2003 Oct 15. 22(20):5323-35.
  PUBMED    14532106
REFERENCE   2  (bases 1 to 8805)
  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"; journalName: "EMBO J. 
            2003 Oct 15. 22(20):5323-35."
COMMENT     Sequence Label: pCMVHA hEZH2
FEATURES             Location/Qualifiers
     source          1..8805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        198..577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        578..781
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     732..752
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     778..802
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          893..1465
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1545..1571
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3924..3943)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3989..4044
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4043..4062)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4402..4418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4402..4418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4415..4437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4426..4442
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4450..4456,4457..4474,4475..4480))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4495..4516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4633..4650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4804..5392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4884..4903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5563..6354,6355..6423))
                     /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     6186..6205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6424..6528)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6596..6614
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6652..6674)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6774..6793
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6987..7009
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7001..7018
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7002..7018
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7429..7574
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             7631..8425
                     /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(7685..7704)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8295..8314
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8619..8638)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8663..8710
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaagc
       61 cctgccatag ccacgggccc cgtgggttag ggacggcgga tcgcggccct ctagagagct
      121 tggcccattg catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc
      181 aacattaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      241 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      301 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      361 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      421 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      481 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      541 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      601 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      661 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      721 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      781 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
      841 aagacaccgg gaccgatcca gcctccggtc gaccgatcct gagaacttca gggtgagttt
      901 ggggaccctt gattgttctt tctttttcgc tattgtaaaa ttcatgttat atggaggggg
      961 caaagttttc agggtgttgt ttagaatggg aagatgtccc ttgtatcacc atggaccctc
     1021 atgataattt tgtttctttc actttctact ctgttgacaa ccattgtctc ctcttatttt
     1081 cttttcattt tctgtaactt tttcgttaaa ctttagcttg catttgtaac gaatttttaa
     1141 attcactttt gtttatttgt cagattgtaa gtactttctc taatcacttt tttttcaagg
     1201 caatcagggt atattatatt gtacttcagc acagttttag agaacaattg ttataattaa
     1261 atgataaggt agaatatttc tgcatataaa ttctggctgg cgtggaaata ttcttattgg
     1321 tagaaacaac tacaccctgg tcatcatcct gcctttctct ttatggttac aatgatatac
     1381 actgtttgag atgaggataa aatactctga gtccaaaccg ggcccctctg ctaaccatgt
     1441 tcatgccttc ttctctttcc tacagctcct gggcaacgtg ctggttgttg tgctgtctca
     1501 tcattttggc aaagaattcc tcgacggatc aattcaccat ggcataccca tacgatgttc
     1561 cagattacgc tagcggatcc atgggccaga ctgggaagaa atctgagaag ggaccagttt
     1621 gttggcggaa gcgtgtaaaa tcagagtaca tgcgactgag acagctcaag aggttcagac
     1681 gagctgatga agtaaagagt atgtttagtt ccaatcgtca gaaaattttg gaaagaacgg
     1741 aaatcttaaa ccaagaatgg aaacagcgaa ggatacagcc tgtgcacatc ctgacttctg
     1801 tgagctcatt gcgcgggact agggagtgtt cggtgaccag tgacttggat tttccaacac
     1861 aagtcatccc attaaagact ctgaatgcag ttgcttcagt acccataatg tattcttggt
     1921 ctcccctaca gcagaatttt atggtggaag atgaaactgt tttacataac attccttata
     1981 tgggagatga agttttagat caggatggta ctttcattga agaactaata aaaaattatg
     2041 atgggaaagt acacggggat agagaatgtg ggtttataaa tgatgaaatt tttgtggagt
     2101 tggtgaatgc ccttggtcaa tataatgatg atgacgatga tgatgatgga gacgatcctg
     2161 aagaaagaga agaaaagcag aaagatctgg aggatcaccg agatgataaa gaaagccgcc
     2221 cacctcggaa atttccttct gataaaattt ttgaagccat ttcctcaatg tttccagata
     2281 agggcacagc agaagaacta aaggaaaaat ataaagaact caccgaacag cagctcccag
     2341 gcgcacttcc tcctgaatgt acccccaaca tagatggacc aaatgctaaa tctgttcaga
     2401 gagagcaaag cttacactcc tttcatacgc ttttctgtag gcgatgtttt aaatatgact
     2461 gcttcctaca tccttttcat gcaacaccca acacttataa gcggaagaac acagaaacag
     2521 ctctagacaa caaaccttgt ggaccacagt gttaccagca tttggaggga gcaaaggagt
     2581 ttgctgctgc tctcaccgct gagcggataa agaccccacc aaaacgtcca ggaggccgca
     2641 gaagaggacg gcttcccaat aacagtagca ggcccagcac ccccaccatt aatgtgctgg
     2701 aatcaaagga tacagacagt gatagggaag cagggactga aacgggggga gagaacaatg
     2761 ataaagaaga agaagagaag aaagatgaaa cttcgagctc ctctgaagca aattctcggt
     2821 gtcaaacacc aataaagatg aagccaaata ttgaacctcc tgagaatgtg gagtggagtg
     2881 gtgctgaagc ctcaatgttt agagtcctca ttggcactta ctatgacaat ttctgtgcca
     2941 ttgctaggtt aattgggacc aaaacatgta gacaggtgta tgagtttaga gtcaaagaat
     3001 ctagcatcat agctccagct cccgctgagg atgtggatac tcctccaagg aaaaagaaga
     3061 ggaaacaccg gttgtgggct gcacactgca gaaagataca gctgaaaaag gacggctcct
     3121 ctaaccatgt ttacaactat caaccctgtg atcatccacg gcagccttgt gacagttcgt
     3181 gcccttgtgt gatagcacaa aatttttgtg aaaagttttg tcaatgtagt tcagagtgtc
     3241 aaaaccgctt tccgggatgc cgctgcaaag cacagtgcaa caccaagcag tgcccgtgct
     3301 acctggctgt ccgagagtgt gaccctgacc tctgtcttac ttgtggagcc gctgaccatt
     3361 gggacagtaa aaatgtgtcc tgcaagaact gcagtattca gcggggctcc aaaaagcatc
     3421 tattgctggc accatctgac gtggcaggct gggggatttt tatcaaagat cctgtgcaga
     3481 aaaatgaatt catctcagaa tactgtggag agattatttc tcaagatgaa gctgacagaa
     3541 gagggaaagt gtatgataaa tacatgtgca gctttctgtt caacttgaac aatgattttg
     3601 tggtggatgc aacccgcaag ggtaacaaaa ttcgttttgc aaatcattcg gtaaatccaa
     3661 actgctatgc aaaagttatg atggttaacg gtgatcacag gataggtatt tttgccaaga
     3721 gagccatcca gactggcgaa gagctgtttt ttgattacag atacagccag gctgatgccc
     3781 tgaagtatgt cggcatcgaa agagaaatgg aaatcccttg aactagttgg atccgtcgag
     3841 gaattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     3901 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
     3961 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     4021 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     4081 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     4141 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     4201 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     4261 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     4321 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacgga
     4381 tcggccgcaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4441 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4501 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4561 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4621 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4681 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4741 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4801 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5221 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5401 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5461 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5521 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5581 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5641 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5701 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5761 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5821 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5881 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5941 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6001 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6061 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6121 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6181 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6241 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6301 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6361 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6421 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6481 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6541 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     6601 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     6661 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     6721 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     6781 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6841 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6901 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6961 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     7021 caagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa gcggaacacg
     7081 tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc
     7141 tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg
     7201 cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg
     7261 ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg
     7321 atctgatggc gcaggggatc aagatcctgc ttcatccccg tggcccgttg ctcgcgtttg
     7381 ctggcggtgt ccccggaaga aatatatttg catgtcttta gttctatgat gacacaaacc
     7441 ccgcccagcg tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt
     7501 cccaggtcca cttcgcatat taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc
     7561 agcgacccgc ttaacagcgt caacagcgtg ccgcagatct gatcaagaga caggatgagg
     7621 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     7681 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     7741 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     7801 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     7861 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     7921 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     7981 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     8041 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     8101 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     8161 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     8221 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     8281 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     8341 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     8401 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     8461 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     8521 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     8581 gagttcttcg cccaccccgg gagatggggg aggctaactg aaacacggaa ggagacaata
     8641 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     8701 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     8761 agaccccatt ggggccaata cgcccgcgtt tcttcctttt cccca
//
