LOCUS       sequence-181559-        8919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mouse Ezh2 with Y637S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8919)
  AUTHORS   Vo BT, Li C, Morgan MA, Theurillat I, Finkelstein D, Wright S, Hyle 
            J, Smith SM, Fan Y, Wang YD, Wu G, Orr BA, Northcott PA, Shilatifard
            A, Sherr CJ, Roussel MF
  TITLE     Inactivation of Ezh2 Upregulates Gfi1 and Drives Aggressive 
            Myc-Driven Group 3 Medulloblastoma.
  JOURNAL   Cell Rep. 2017 Mar 21;18(12):2907-2917. doi: 
            10.1016/j.celrep.2017.02.073.
  PUBMED    28329683
REFERENCE   2  (bases 1 to 8919)
  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: 
            "10.1016/j.celrep.2017.02"; journalName: "Cell Rep"; date: 
            "2017-03-21- 21"; volume: "18"; issue: "12"; pages: "2907-2917"
COMMENT     Sequence Label: MSCV-mouse Ezh2 Y637S-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..58
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             258..774
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    838..1179
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1246..1662
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1592..1614
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1630..1646
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4138..4687
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4282..4299)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4509..4528
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4690..4692,4693..4695,4696..5409)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4690..4692,4693..4695,4696..5406)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4735..4756)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4996..5015)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5343..5364
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5571..6085
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6247..6263
                     /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(6271..6277,6278..6295,6296..6301))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6316..6337
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6454..6471)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6625..7213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6705..6724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7384..8175,8176..8244))
                     /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     8007..8026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8245..8349)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8417..8435
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8473..8495)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8595..8614
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8808..8830
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
       61 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggcga
      121 ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt gactcaacaa
      181 tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt
      241 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
      301 acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt
      361 taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg
      421 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct
      481 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
      541 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
      601 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg
      661 ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg actcgctgat
      721 ccttgggagg gtctcctcag attgattgac tgcccacctc gggggtcttt catttggagg
      781 ttccaccgag atttggagac ccctgcccag ggaccaccga cccccccgcc gggaggtaag
      841 ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc cggcatctaa
      901 tgtttgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg gacccgtggt
      961 ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg
     1021 ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga
     1081 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
     1141 ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg
     1201 tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc
     1261 actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag
     1321 tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt
     1381 aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc
     1441 aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg
     1501 gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg
     1561 cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct
     1621 cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattgg gatccactag
     1681 taacggccgc cagtgtgctg gaattcagga tcgtcgaccc acgcgtccgc ggacgcgtgg
     1741 gttgggggcg acgcgcggga gaggcgcggg ctggcgcgcg ggacgaagaa taatcatggg
     1801 ccagactggg aagaaatctg agaagggacc ggtttgttgg cggaagcgtg taaaatcaga
     1861 gtacatgaga ctgagacagc tcaagaggtt cagaagagct gatgaagtaa agactatgtt
     1921 tagttccaat cgtcagaaaa ttttggaaag aactgaaacc ttaaaccaag agtggaagca
     1981 gcggaggata cagcctgtgc acatcatgac ttctgtgagc tcattgcgcg ggactaggga
     2041 gtgttcagtc accagtgact tggattttcc agcacaagtc atcccgttaa agaccctgaa
     2101 tgcagtcgcc tcggtgccta taatgtactc ttggtcgccc ttacaacaga attttatggt
     2161 ggaagacgaa actgtcttac ataacattcc ttacatgggg gatgaagttc tggatcagga
     2221 cggcactttc attgaagaac taataaaaaa ttatgatgga aaagtgcatg gtgacagaga
     2281 atgtggattt ataaatgatg aaatttttgt ggagttggta aatgctcttg gtcaatataa
     2341 tgatgatgat gatgacgatg atggagatga tccagatgaa agagaagaaa aacagaaaga
     2401 tctagaggat aatcgagatg ataaagaaac ttgcccacct cggaaatttc ctgctgataa
     2461 aatatttgaa gccatttcct caatgtttcc agataagggc accgcagaag aactgaagga
     2521 aaaatataaa gaactcacgg agcagcagct cccaggtgct ctgcctcctg aatgtactcc
     2581 aaacatcgat ggaccaaatg ccaaatctgt tcagagggag caaagcttgc attcatttca
     2641 tacgctcttc tgtcgacgat gttttaagta tgactgcttc ctacatccct tccatgcaac
     2701 acccaacaca tataagagga agaacacaga aacagctttg gacaacaagc cttgtggacc
     2761 acagtgttac cagcatctgg agggagctaa ggagtttgct gctgctctta ctgctgagcg
     2821 gataaagaca ccacctaaac gcccaggggg ccgcagaaga ggaagacttc cgaataacag
     2881 tagcagaccc agcaccccca ccatcagtgt gctggagtca aaggatacag acagtgacag
     2941 agaagcaggg actgaaactg ggggagagaa caacgataaa gaagaagaag agaaaaaaga
     3001 tgagacgtcc agctcctctg aagcaaattc tcggtgtcaa acaccaataa agatgaagcc
     3061 aaatattgaa cctcctgaga atgtggagtg gagtggtgct gaagcctcca tgtttagagt
     3121 cctcattggt acttactacg ataacttttg tgccattgct aggctaattg ggaccaaaac
     3181 atgtagacag gtgtatgagt ttagagtcaa ggagtccagt atcatagcac ctgttcccac
     3241 tgaggatgta gacactcctc caagaaagaa gaaaaggaaa catcggttgt gggctgcaca
     3301 ctgcagaaag atacaactga aaaaggacgg ctcctctaac catgtttaca actatcaacc
     3361 ctgtgaccat ccacggcagc cttgtgacag ttcgtgccct tgtgtgatag cacaaaattt
     3421 ttgtgaaaag ttttgtcaat gtagttcaga gtgtcaaaac cgctttcctg gatgtcggtg
     3481 caaagcacaa tgcaacacca aacagtgtcc atgctacctg gctgtccgag agtgtgaccc
     3541 tgacctctgt ctcacgtgtg gagctgctga ccattgggac agtaaaaatg tatcctgtaa
     3601 gaactgtagc attcagcggg gctctaaaaa gcacttactg ctggcaccgt ctgatgtggc
     3661 aggctggggc atctttatca aagatcctgt acagaaaaat gaattcatct cagaaagctg
     3721 tggggagatt atttctcagg atgaagcaga cagaagagga aaagtgtatg acaaatacat
     3781 gtgcagcttt ctgttcaact tgaacaatga ttttgtggtg gatgcaaccc gaaagggcaa
     3841 caaaattcgt tttgctaatc attcagtaaa tccaaactgc tatgcaaaag ttatgatggt
     3901 taatggtgac cacaggatag gcatctttgc taagagggct atccagactg gtgaagagtt
     3961 gttttttgat tacagataca gccaggctga tgccctgaag tatgtgggca tcgaacgaga
     4021 aatggaaatc ccttgacatc tactacctct tccccttctc tctgaaacag cctgaattct
     4081 gcagatatcc atcacactgg cggccgctcg agcgggatca attccgcccc ccccctaacg
     4141 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     4201 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     4261 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     4321 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     4381 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     4441 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     4501 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     4561 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     4621 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     4681 gataatacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     4741 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4801 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     4861 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     4921 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4981 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     5041 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     5101 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     5161 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     5221 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     5281 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     5341 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     5401 tacaagtaaa gcggccgcga ctctagagtc gacctgcagg catgcaagct tcaggtagcc
     5461 ggctaacgtt aacaaccggt acctctagaa ctatagctag catgcgcaaa tttaaagcgc
     5521 tgatatcgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     5581 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac
     5641 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg
     5701 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     5761 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     5821 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     5881 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     5941 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt
     6001 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     6061 ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc
     6121 tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac
     6181 atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt
     6241 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6301 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6361 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6421 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6481 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6541 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6601 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6661 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6721 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6781 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6841 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6901 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6961 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7021 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7081 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7141 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7201 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7261 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7321 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7381 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7441 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7501 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7561 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7621 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7681 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7741 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7801 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7861 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7921 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7981 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8041 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8101 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8161 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8221 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8281 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8341 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     8401 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     8461 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     8521 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     8581 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     8641 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     8701 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     8761 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     8821 ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     8881 gccacggggc ctgccaccat acccacgccg aaacaagcg
//
