LOCUS       sequence-207627-        8919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mouse Ezh2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8919)
  TITLE     Roussel lab unpublished plasmids
  JOURNAL   Unpublished
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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: MSCV-mouse Ezh2-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     88..107
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             307..823
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    887..1228
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1295..1711
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1641..1663
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1679..1695
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4187..4736
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4331..4348)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4558..4577
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4739..4741,4742..4744,4745..5458)
                     /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(4739..4741,4742..4744,4745..5455)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4784..4805)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5045..5064)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5392..5413
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5620..6134
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6296..6312
                     /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(6320..6326,6327..6344,6345..6350))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6365..6386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6503..6520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6674..7262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6754..6773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7433..8224,8225..8293))
                     /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     8056..8075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8294..8398)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8466..8484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8522..8544)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8644..8663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8857..8879
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
       61 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
      121 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt
      181 tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct
      241 gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa
      301 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
      361 caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag
      421 agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
      481 ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc
      541 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
      601 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
      661 cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtat
      721 tcccaataaa gcctcttgct gtttgcatcc gaatcgtgga ctcgctgatc cttgggaggg
      781 tctcctcaga ttgattgact gcccacctcg ggggtctttc atttggaggt tccaccgaga
      841 tttggagacc cctgcccagg gaccaccgac ccccccgccg ggaggtaagc tggccagcgg
      901 tcgtttcgtg tctgtctctg tctttgtgcg tgtttgtgcc ggcatctaat gtttgcgcct
      961 gcgtctgtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
     1021 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
     1081 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
     1141 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
     1201 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
     1261 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
     1321 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1381 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1441 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1501 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1561 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1621 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1681 ttatccagcc ctcactcctt ctctaggcgc cggaattggg atccactagt aacggccgcc
     1741 agtgtgctgg aattcaggat cgtcgaccca cgcgtccgcg gacgcgtggg ttgggggcga
     1801 cgcgcgggag aggcgcgggc tggcgcgcgg gacgaagaat aatcatgggc cagactggga
     1861 agaaatctga gaagggaccg gtttgttggc ggaagcgtgt aaaatcagag tacatgagac
     1921 tgagacagct caagaggttc agaagagctg atgaagtaaa gactatgttt agttccaatc
     1981 gtcagaaaat tttggaaaga actgaaacct taaaccaaga gtggaagcag cggaggatac
     2041 agcctgtgca catcatgact tctgtgagct cattgcgcgg gactagggag tgttcagtca
     2101 ccagtgactt ggattttcca gcacaagtca tcccgttaaa gaccctgaat gcagtcgcct
     2161 cggtgcctat aatgtactct tggtcgccct tacaacagaa ttttatggtg gaagacgaaa
     2221 ctgtcttaca taacattcct tacatggggg atgaagttct ggatcaggac ggcactttca
     2281 ttgaagaact aataaaaaat tatgatggaa aagtgcatgg tgacagagaa tgtggattta
     2341 taaatgatga aatttttgtg gagttggtaa atgctcttgg tcaatataat gatgatgatg
     2401 atgacgatga tggagatgat ccagatgaaa gagaagaaaa acagaaagat ctagaggata
     2461 atcgagatga taaagaaact tgcccacctc ggaaatttcc tgctgataaa atatttgaag
     2521 ccatttcctc aatgtttcca gataagggca ccgcagaaga actgaaggaa aaatataaag
     2581 aactcacgga gcagcagctc ccaggtgctc tgcctcctga atgtactcca aacatcgatg
     2641 gaccaaatgc caaatctgtt cagagggagc aaagcttgca ttcatttcat acgctcttct
     2701 gtcgacgatg ttttaagtat gactgcttcc tacatccctt ccatgcaaca cccaacacat
     2761 ataagaggaa gaacacagaa acagctttgg acaacaagcc ttgtggacca cagtgttacc
     2821 agcatctgga gggagctaag gagtttgctg ctgctcttac tgctgagcgg ataaagacac
     2881 cacctaaacg cccagggggc cgcagaagag gaagacttcc gaataacagt agcagaccca
     2941 gcacccccac catcagtgtg ctggagtcaa aggatacaga cagtgacaga gaagcaggga
     3001 ctgaaactgg gggagagaac aacgataaag aagaagaaga gaaaaaagat gagacgtcca
     3061 gctcctctga agcaaattct cggtgtcaaa caccaataaa gatgaagcca aatattgaac
     3121 ctcctgagaa tgtggagtgg agtggtgctg aagcctccat gtttagagtc ctcattggta
     3181 cttactacga taacttttgt gccattgcta ggctaattgg gaccaaaaca tgtagacagg
     3241 tgtatgagtt tagagtcaag gagtccagta tcatagcacc tgttcccact gaggatgtag
     3301 acactcctcc aagaaagaag aaaaggaaac atcggttgtg ggctgcacac tgcagaaaga
     3361 tacaactgaa aaaggacggc tcctctaacc atgtttacaa ctatcaaccc tgtgaccatc
     3421 cacggcagcc ttgtgacagt tcgtgccctt gtgtgatagc acaaaatttt tgtgaaaagt
     3481 tttgtcaatg tagttcagag tgtcaaaacc gctttcctgg atgtcggtgc aaagcacaat
     3541 gcaacaccaa acagtgtcca tgctacctgg ctgtccgaga gtgtgaccct gacctctgtc
     3601 tcacgtgtgg agctgctgac cattgggaca gtaaaaatgt atcctgtaag aactgtagca
     3661 ttcagcgggg ctctaaaaag cacttactgc tggcaccgtc tgatgtggca ggctggggca
     3721 tctttatcaa agatcctgta caaaaaaatg aattcatctc agaatactgt ggggagatta
     3781 tttctcagga tgaagcagac agaagaggaa aagtgtatga caaatacatg tgcagctttc
     3841 tgttcaactt gaacaatgat tttgtggtgg atgcaacccg aaagggcaac aaaattcgtt
     3901 ttgctaatca ttcagtaaat ccaaactgct atgcaaaagt tatgatggtt aatggtgacc
     3961 acaggatagg catctttgct aagagggcta tccagactgg tgaagagttg ttttttgatt
     4021 acagatacag ccaggctgat gccctgaagt atgtgggcat cgaacgagaa atggaaatcc
     4081 cttgacatct actacctctt ccccttctct ctgaaacagc ctgaattctg cagatatcca
     4141 tcacactggc ggccgctcga gcgggatcaa ttccgccccc cccctaacgt tactggccga
     4201 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     4261 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     4321 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     4381 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     4441 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     4501 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     4561 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     4621 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     4681 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg ataataccat
     4741 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     4801 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     4861 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     4921 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     4981 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     5041 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     5101 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     5161 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     5221 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     5281 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     5341 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     5401 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     5461 cggccgcgac tctagagtcg acctgcaggc atgcaagctt caggtagccg gctaacgtta
     5521 acaaccggta cctctagaac tatagctagc atgcgcaaat ttaaagcgct gatatcgata
     5581 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     5641 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa
     5701 tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat
     5761 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     5821 gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     5881 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     5941 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     6001 tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     6061 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     6121 cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga
     6181 gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata
     6241 ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt
     6301 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6361 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6421 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6481 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6541 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6601 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6661 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6721 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6781 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6841 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6901 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6961 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7021 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7081 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7141 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7201 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7261 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7321 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7381 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7441 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7501 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7561 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7621 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7681 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7741 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7801 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7861 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7921 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7981 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8041 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8101 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8161 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8221 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8281 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8341 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8401 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     8461 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     8521 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     8581 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     8641 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     8701 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     8761 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     8821 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     8881 cggcgcaagg aatggtgcat gcaaggagat ggcgcccaa
//
