LOCUS       sequence-244370-        8362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mouse Ezh2 with a SET domain deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8362)
  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 8362)
  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 delta SETdomain-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1420..1429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1429..1452
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3323..3872
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3467..3484)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3694..3713
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3875..3877,3878..3880,3881..4594)
                     /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(3875..3877,3878..3880,3881..4591)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3920..3941)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4181..4200)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4528..4549
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4756..5270
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5432..5448
                     /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(5456..5462,5463..5480,5481..5486))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5501..5522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5639..5656)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5810..6398)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5890..5909)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6569..7360,7361..7429))
                     /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     7192..7211
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7430..7534)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7602..7620
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7658..7680)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7780..7799
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7993..8015
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8143..8162
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatcctgg ccaccatgga ctacaaggac
     1441 gacgatgaca agatgggcca gactgggaag aaatctgaga agggaccggt ttgttggcgg
     1501 aagcgtgtaa aatcagagta catgagactg agacagctca agaggttcag aagagctgat
     1561 gaagtaaaga ctatgtttag ttccaatcgt cagaaaattt tggaaagaac tgaaacctta
     1621 aaccaagagt ggaagcagcg gaggatacag cctgtgcaca tcatgacttc tgtgagctca
     1681 ttgcgcggga ctagggagtg ttcagtcacc agtgacttgg attttccagc acaagtcatc
     1741 ccgttaaaga ccctgaatgc agtcgcctcg gtgcctataa tgtactcttg gtcgccctta
     1801 caacagaatt ttatggtgga agacgaaact gtcttacata acattcctta catgggggat
     1861 gaagttctgg atcaggacgg cactttcatt gaagaactaa taaaaaatta tgatggaaaa
     1921 gtgcatggtg acagagaatg tggatttata aatgatgaaa tttttgtgga gttggtaaat
     1981 gctcttggtc aatataatga tgatgatgat gacgatgatg gagatgatcc agatgaaaga
     2041 gaagaaaaac agaaagatct agaggataat cgagatgata aagaaacttg cccacctcgg
     2101 aaatttcctg ctgataaaat atttgaagcc atttcctcaa tgtttccaga taagggcacc
     2161 gcagaagaac tgaaggaaaa atataaagaa ctcacggagc agcagctccc aggtgctctg
     2221 cctcctgaat gtactccaaa catcgatgga ccaaatgcca aatctgttca gagggagcaa
     2281 agcttgcatt catttcatac gctcttctgt cgacgatgtt ttaagtatga ctgcttccta
     2341 catcccttcc atgcaacacc caacacatat aagaggaaga acacagaaac agctttggac
     2401 aacaagcctt gtggaccaca gtgttaccag catctggagg gagctaagga gtttgctgct
     2461 gctcttactg ctgagcggat aaagacacca cctaaacgcc cagggggccg cagaagagga
     2521 agacttccga ataacagtag cagacccagc acccccacca tcagtgtgct ggagtcaaag
     2581 gatacagaca gtgacagaga agcagggact gaaactgggg gagagaacaa cgataaagaa
     2641 gaagaagaga aaaaagatga gacgtccagc tcctctgaag caaattctcg gtgtcaaaca
     2701 ccaataaaga tgaagccaaa tattgaacct cctgagaatg tggagtggag tggtgctgaa
     2761 gcctccatgt ttagagtcct cattggtact tactacgata acttttgtgc cattgctagg
     2821 ctaattggga ccaaaacatg tagacaggtg tatgagttta gagtcaagga gtccagtatc
     2881 atagcacctg ttcccactga ggatgtagac actcctccaa gaaagaagaa aaggaaacat
     2941 cggttgtggg ctgcacactg cagaaagata caactgaaaa aggacggctc ctctaaccat
     3001 gtttacaact atcaaccctg tgaccatcca cggcagcctt gtgacagttc gtgcccttgt
     3061 gtgatagcac aaaatttttg tgaaaagttt tgtcaatgta gttcagagtg tcaaaaccgc
     3121 tttcctggat gtcggtgcaa agcacaatgc aacaccaaac agtgtccatg ctacctggct
     3181 gtccgagagt gtgaccctga cctctgtctc acgtgtggag ctgctgacca ttgggacagt
     3241 aaaaatgtat cctgtaagaa ctgtagcatt cagcggggct ctaaaaagtg actcgagcgg
     3301 gatcaattcc gccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     3361 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     3421 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     3481 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     3541 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     3601 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     3661 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     3721 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     3781 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     3841 gacgtggttt tcctttgaaa aacacgataa taccatggtg agcaagggcg aggagctgtt
     3901 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3961 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     4021 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     4081 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4141 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     4201 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     4261 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     4321 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     4381 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4441 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     4501 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4561 gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gagtcgacct
     4621 gcaggcatgc aagcttcagg tagccggcta acgttaacaa ccggtacctc tagaactata
     4681 gctagcatgc gcaaatttaa agcgctgata tcgataaaat aaaagatttt atttagtctc
     4741 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     4801 ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag
     4861 gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     4921 cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga
     4981 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     5041 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     5101 aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt
     5161 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     5221 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt
     5281 ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact
     5341 caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt cattatggac
     5401 agcgcagaaa gagctgggga gaattgtgaa attgttatcc gctcacaatt ccacacaaca
     5461 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5521 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5581 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5641 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5701 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5761 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5821 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5881 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5941 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6001 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6061 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6121 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6181 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6241 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6301 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6361 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6421 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6481 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6541 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6601 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6661 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6721 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6781 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6841 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6901 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6961 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7021 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7081 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7141 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7201 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7261 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7321 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7381 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7441 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7501 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7561 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     7621 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     7681 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     7741 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     7801 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     7861 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     7921 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     7981 cgccagggtt ttcccagtca cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa
     8041 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     8101 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     8161 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     8221 gcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag ttttgactca
     8281 acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa agattttatt
     8341 tagtctccag aaaaaggggg ga
//
