LOCUS       sequence-203895-        8800 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 8800)
  TITLE     Roussel lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8800)
  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            "Unpublished"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..8800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    3755..4303
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3899..3916)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4126..4145
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4312..4314,4315..4317,4318..5031)
                     /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(4312..4314,4315..4317,4318..5028)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4357..4378)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4618..4637)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4965..4986
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5193..5707
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5869..5885
                     /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(5893..5899,5900..5917,5918..5923))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5938..5959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6076..6093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6247..6835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6327..6346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7006..7797,7798..7866))
                     /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     7629..7648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7867..7971)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8039..8057
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8095..8117)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8217..8236
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8430..8452
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8580..8599
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8799..8800,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaattg ggatcccgga cgcgtgggtt gggggcgacg
     1441 cgcgggagag gcgcgggctg gcgcgcggga cgaagaataa tcatgggcca gactgggaag
     1501 aaatctgaga agggaccggt ttgttggcgg aagcgtgtaa aatcagagta catgagactg
     1561 agacagctca agaggttcag aagagctgat gaagtaaaga ctatgtttag ttccaatcgt
     1621 cagaaaattt tggaaagaac tgaaacctta aaccaagagt ggaagcagcg gaggatacag
     1681 cctgtgcaca tcatgacttc tgtgagctca ttgcgcggga ctagggagtg ttcagtcacc
     1741 agtgacttgg attttccagc acaagtcatc ccgttaaaga ccctgaatgc agtcgcctcg
     1801 gtgcctataa tgtactcttg gtcgccctta caacagaatt ttatggtgga agacgaaact
     1861 gtcttacata acattcctta catgggggat gaagttctgg atcaggacgg cactttcatt
     1921 gaagaactaa taaaaaatta tgatggaaaa gtgcatggtg acagagaatg tggatttata
     1981 aatgatgaaa tttttgtgga gttggtaaat gctcttggtc aatataatga tgatgatgat
     2041 gacgatgatg gagatgatcc agatgaaaga gaagaaaaac agaaagatct agaggataat
     2101 cgagatgata aagaaacttg cccacctcgg aaatttcctg ctgataaaat atttgaagcc
     2161 atttcctcaa tgtttccaga taagggcacc gcagaagaac tgaaggaaaa atataaagaa
     2221 ctcacggagc agcagctccc aggtgctctg cctcctgaat gtactccaaa catcgatgga
     2281 ccaaatgcca aatctgttca gagggagcaa agcttgcatt catttcatac gctcttctgt
     2341 cgacgatgtt ttaagtatga ctgcttccta catcccttcc atgcaacacc caacacatat
     2401 aagaggaaga acacagaaac agctttggac aacaagcctt gtggaccaca gtgttaccag
     2461 catctggagg gagctaagga gtttgctgct gctcttactg ctgagcggat aaagacacca
     2521 cctaaacgcc cagggggccg cagaagagga agacttccga ataacagtag cagacccagc
     2581 acccccacca tcagtgtgct ggagtcaaag gatacagaca gtgacagaga agcagggact
     2641 gaaactgggg gagagaacaa cgataaagaa gaagaagaga aaaaagatga gacgtccagc
     2701 tcctctgaag caaattctcg gtgtcaaaca ccaataaaga tgaagccaaa tattgaacct
     2761 cctgagaatg tggagtggag tggtgctgaa gcctccatgt ttagagtcct cattggtact
     2821 tactacgata acttttgtgc cattgctagg ctaattggga ccaaaacatg tagacaggtg
     2881 tatgagttta gagtcaagga gtccagtatc atagcacctg ttcccactga ggatgtagac
     2941 actcctccaa gaaagaagaa aaggaaacat cggttgtggg ctgcacactg cagaaagata
     3001 caactgaaaa aggacggctc ctctaaccat gtttacaact atcaaccctg tgaccatcca
     3061 cggcagcctt gtgacagttc gtgcccttgt gtgatagcac aaaatttttg tgaaaagttt
     3121 tgtcaatgta gttcagagtg tcaaaaccgc tttcctggat gtcggtgcaa agcacaatgc
     3181 aacaccaaac agtgtccatg ctacctggct gtccgagagt gtgaccctga cctctgtctc
     3241 acgtgtggag ctgctgacca ttgggacagt aaaaatgtat cctgtaagaa ctgtagcatt
     3301 cagcggggct ctaaaaagca cttactgctg gcaccgtctg atgtggcagg ctggggcatc
     3361 tttatcaaag atcctgtaca aaaaaatgaa ttcatctcag aatactgtgg ggagattatt
     3421 tctcaggatg aagcagacag aagaggaaaa gtgtatgaca aatacatgtg cagctttctg
     3481 ttcaacttga acaatgattt tgtggtggat gcaacccgaa agggcaacaa aattcgtttt
     3541 gctaatcatt cagtaaatcc aaactgctat gcaaaagtta tgatggttaa tggtgaccac
     3601 aggataggca tctttgctaa gagggctatc cagactggtg aagagttgtt ttttgattac
     3661 agatacagcc aggctgatgc cctgaagtat gtgggcatcg aacgagaaat ggaaatccct
     3721 tgactcgagc gggatcaatt ccgccccccc cctaacgtta ctggccgaag ccgcttggaa
     3781 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     3841 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     3901 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     3961 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     4021 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     4081 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     4141 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     4201 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc
     4261 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata atacctgtac aatggtgagc
     4321 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     4381 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     4441 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     4501 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     4561 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     4621 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     4681 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     4741 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     4801 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     4861 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     4921 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     4981 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc
     5041 gactctagag tcgacctgca ggcatgcaag cttcaggtag ccggctaacg ttaacaaccg
     5101 gtacctctag aactatagct agcatgcgca aatttaaagc gctgatatcg ataaaataaa
     5161 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     5221 agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa
     5281 gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg
     5341 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc
     5401 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     5461 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     5521 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata
     5581 gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     5641 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     5701 ctttcatggg taacagtttc ttgaagttgg agaacaacat tctgagggta ggagtcgaat
     5761 attaagtaat cctgactcaa ttagccactg ttttgaatcc acatactcca atactcctga
     5821 aatagttcat tatggacagc gcagaaagag ctggggagaa ttgtgaaatt gttatccgct
     5881 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5941 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6001 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6061 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6121 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6181 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6421 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6661 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6781 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6841 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6901 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6961 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7021 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7081 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7141 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7201 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7261 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7321 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7381 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7441 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7501 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7561 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7621 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7681 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7741 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7801 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7861 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7921 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7981 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     8041 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     8101 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     8161 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     8221 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     8281 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     8341 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     8401 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggcgca
     8461 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     8521 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     8581 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     8641 atgcgtccgg cgtagaggcg attagtccaa tttgttaaag acaggatatc agtggtccag
     8701 gctctagttt tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata
     8761 aaataaaaga ttttatttag tctccagaaa aaggggggaa
//
