LOCUS       sequence-138257-        7883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human EED with HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7883)
  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 7883)
  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 EED wt
FEATURES             Location/Qualifiers
     source          1..7883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     607..631
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          722..1294
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1374..1400
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2831..2850)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2896..2951
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2950..2969)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3309..3325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3309..3325)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3322..3344)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3333..3349
                     /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(3357..3363,3364..3381,3382..3387))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3402..3423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3540..3557)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3711..4299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3791..3810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4470..5261,5262..5330))
                     /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     5093..5112
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5331..5435)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5503..5521
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5559..5581)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5681..5700
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5894..5916
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5908..5925
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5909..5925
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6336..6481
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6538..7332
                     /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(6592..6611)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7202..7221
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7526..7545)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7570..7617
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
      661 cctccataga agacaccggg accgatccag cctccggtcg accgatcctg agaacttcag
      721 ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat tcatgttata
      781 tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct tgtatcacca
      841 tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac cattgtctcc
      901 tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc atttgtaacg
      961 aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct aatcactttt
     1021 ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga gaacaattgt
     1081 tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc gtggaaatat
     1141 tcttattggt agaaacaact acaccctggt catcatcctg cctttctctt tatggttaca
     1201 atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg gcccctctgc
     1261 taaccatgtt catgccttct tctctttcct acagctcctg ggcaacgtgc tggttgttgt
     1321 gctgtctcat cattttggca aagaattcct cgacggatca attcaccatg gcatacccat
     1381 acgatgttcc agattacgct agcggatcca tgtccgagag ggaagtgtcg actgcgccgg
     1441 cgggaacaga catgcccgcg gccaagaagc agaagttgag cagcgacgag aacagcaacc
     1501 cggacctctc gggagacgaa aatgacgatg ctgtcagtat tgagagtggc acaaacacag
     1561 aacgcccgga cacgcccaca aatacgccaa atgcaccagg aaggaaaagc tggggaaagg
     1621 gaaaatggaa gtcaaagaaa tgcaaatatt ctttcaaatg tgtgaacagc ctcaaggaag
     1681 atcataacca gccattgttt ggagttcagt ttaactggca cagtaaagaa ggagaccctc
     1741 tggtgtttgc aactgtggga agcaacagag taaccttata cgaatgccat tcacaggggg
     1801 agatacggtt attgcagtcc tatgtagatg ctgatgcaga tgaaaacttt tacacttgtg
     1861 catggaccta tgatagcaac accagccacc ctctattagc agttgctgga tctagaggca
     1921 ttataagaat aattaatcct ataacaatgc agtgtataaa gcactatgtt ggccatggaa
     1981 atgctatcaa tgagctgaaa ttccacccac gagacccaaa ccttctcctg tcagtaagta
     2041 aagatcatgc tttacggtta tggaatatcc aaacagacac tcttgtggca atattcggag
     2101 gtgtggaagg gcacagagat gaagttctga gtgctgatta tgatcttttg ggtgaaaaaa
     2161 taatgtcctg tggtatggat cactctctta aactgtggag aatcaactca aagaggatga
     2221 tgaatgcaat taaggagtct tatgattata acccaaacaa aactaacagg ccatttattt
     2281 cccagaaaat ccactttcct gacttttcta ccagagacat acataggaat tatgttgatt
     2341 gtgtgcgatg gttaggcgat ttgatacttt ccaagtcttg tgaaaatgcc attgtatgct
     2401 ggaaacctgg caaaatggag gatgatatag ataaaattaa acctagtgag tctaatgtga
     2461 ctattcttgg gcgatttgat tacagccagt gtgacatttg gtacatgagg ttttctatgg
     2521 atttctggca aaagatgctt gcattgggca atcaggttgg caaactgtat gtttgggatt
     2581 tagaagtaga agatcctcat aaagccaaat gcacaacact gacccatcat aaatgtggcg
     2641 cggctattcg acaaaccagt ttcagtaggg atagcagcat cctcatagct gtctgcgatg
     2701 atgccagcat ttggcgctgg gatcgacttc gataaggatc cgtcgaggaa ttcactcctc
     2761 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     2821 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     2881 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     2941 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     3001 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     3061 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     3121 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     3181 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     3241 tcccagtcat agctgtccct cttctcttat ggagatccct cgacggatcg gccgcaattc
     3301 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3361 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3421 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3481 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3541 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3601 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3661 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3721 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3781 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3841 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3901 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3961 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4021 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4081 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4141 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4201 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4261 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4321 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4381 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4441 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4501 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4561 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4621 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4681 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4741 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4801 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4861 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4921 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4981 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5041 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5101 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5161 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5221 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5281 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5341 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5401 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5461 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5521 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5581 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5641 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     5701 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     5761 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5821 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     5881 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttcacgct
     5941 gccgcaagca ctcagggcgc aagggctgct aaaggaagcg gaacacgtag aaagccagtc
     6001 cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa
     6061 acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact
     6121 gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     6181 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     6241 ggggatcaag atcctgcttc atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc
     6301 cggaagaaat atatttgcat gtctttagtt ctatgatgac acaaaccccg cccagcgtct
     6361 tgtcattggc gaattcgaac acgcagatgc agtcggggcg gcgcggtccc aggtccactt
     6421 cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc gacccgctta
     6481 acagcgtcaa cagcgtgccg cagatctgat caagagacag gatgaggatc gtttcgcatg
     6541 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     6601 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6661 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     6721 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     6781 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     6841 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     6901 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     6961 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     7021 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     7081 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     7141 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     7201 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     7261 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     7321 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     7381 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     7441 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     7501 accccgggag atgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     7561 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     7621 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     7681 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     7741 cagggctcgc agccaacgtc ggggcggcaa gccctgccat agccacgggc cccgtgggtt
     7801 agggacggcg gatcgcggcc ctctagagag cttggcccat tgcatacgtt gtatccatat
     7861 cataatatgt acatttatat tgg
//
