LOCUS       sequence-358093-        7970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Role of Mdm2 splicing variants in tumorigenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7970)
  AUTHORS   Dang J, Kuo ML, Eischen CM, Stepanova L, Sherr CJ, Roussel MF
  TITLE     The RING domain of Mdm2 can inhibit cell proliferation.
  JOURNAL   Cancer Res. 2002 Feb 15;62(4):1222-30.
  PUBMED    11861407
REFERENCE   2  (bases 1 to 7970)
  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: "Cancer 
            Res."; date: "2002-02-15"; volume: "62(4)"; pages: "1222-30"
COMMENT     Sequence Label: MSCV--Mouse Mdm2 wt-IRES-CFP
FEATURES             Location/Qualifiers
     source          1..7970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             200..716
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    780..1121
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1188..1604
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1534..1556
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1572..1588
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    3131..3680
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3275..3292)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3502..3521
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3683..3685,3686..3688,3689..4402)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3728..3749)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3989..4008)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4336..4357
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4564..5078
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5240..5256
                     /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(5264..5270,5271..5288,5289..5294))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5309..5330
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5447..5464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5618..6206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5698..5717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6377..7168,7169..7237))
                     /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     7000..7019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7238..7342)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7410..7428
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7466..7488)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7588..7607
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7801..7823
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7951..7970
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc
       61 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac
      121 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta
      181 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
      241 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
      301 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
      361 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
      421 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
      481 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
      541 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
      601 cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggactcgctg
      661 atccttggga gggtctcctc agattgattg actgcccacc tcgggggtct ttcatttgga
      721 ggttccaccg agatttggag acccctgccc agggaccacc gacccccccg ccgggaggta
      781 agctggccag cggtcgtttc gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct
      841 aatgtttgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg
      901 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
      961 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
     1021 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
     1081 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
     1141 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     1201 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     1261 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     1321 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     1381 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     1441 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
     1501 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
     1561 ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggaatt caggccaatg
     1621 tgcaatacca acatgtctgt gtctaccgag ggtgctgcaa gcacctcaca gattccagct
     1681 tcggaacaag agactctggt tagaccaaaa ccattgcttt tgaagttgtt aaagtccgtt
     1741 ggagcgcaaa acgacactta cactatgaaa gagattatat tttatattgg ccagtatatt
     1801 atgactaaga ggttatatga cgagaagcag cagcacattg tgtattgttc aaatgatctc
     1861 ctaggagatg tgtttggagt cccgagtttc tctgtgaagg agcacaggaa aatatatgca
     1921 atgatctaca gaaatttagt ggctgtaagt cagcaagact ctggcacatc gctgagtgag
     1981 agcagacgtc agcctgaagg tgggagtgat ctgaaggatc ctttgcaagc gccaccagaa
     2041 gagaaacctt catcttctga tttaatttct agactgtcta cctcatctag aaggagatcc
     2101 attagtgaga cagaagagaa cacagatgag ctacctgggg agcggcaccg gaagcgccgc
     2161 aggtccctgt cctttgatcc gagcctgggt ctgtgtgagc tgagggagat gtgcagcggc
     2221 ggcagcagca gcagtagcag cagcagcagc gagtccacag agacgccctc gcatcaggat
     2281 cttgacgatg gcgtaagtga gcattctggt gattgcctgg atcaggattc agtttctgat
     2341 cagtttagcg tggaatttga agttgagtct ctggactcgg aagattacag cctgagtgac
     2401 gaagggcacg agctctcaga tgaggatgat gaggtctatc gggtcacagt ctatcagaca
     2461 ggagaaagcg atacagactc ttttgaaggg gatcctgaga tttccttagc tgactattgg
     2521 aagtgtacct catgcaatga aatgaatcct ccccttccat cacactgcaa aagatgctgg
     2581 acccttcgtg agaactggct tccagacgat aaggggaaag ataaagtgga aatctctgaa
     2641 aaagccaaac tggaaaactc agctcaggca gaagaaggct tggatgtgcc tgatggcaaa
     2701 aagctgacag agaatgatgc taaagagcca tgtgctgagg aggacagcga ggagaaggcc
     2761 gaacagacgc ccctgtccca ggagagtgac gactattccc aaccatcgac ttccagcagc
     2821 attgtttata gcagccaaga aagcgtgaaa gagttgaagg aggaaacgca ggacaaagac
     2881 gagagtgtgg aatctagctt ctccctgaat gccatcgaac catgtgtgat ctgccagggg
     2941 cggcctaaaa atggctgcat tgttcacggc aagactggac acctcatgtc atgtttcacg
     3001 tgtgcaaaga agctaaaaaa aagaaacaag ccctgcccag tgtgcagaca gccaatccaa
     3061 atgattgtgc taacttactt caactagctg acctgctcac tcgagcggga tcaattccgc
     3121 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     3181 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3241 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3301 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     3361 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3421 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3481 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3541 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     3601 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3661 ctttgaaaaa cacgataata ccatggtgag caagggcgag gagctgttca ccggggtggt
     3721 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3781 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3841 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag
     3901 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3961 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     4021 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     4081 ggacggcaac atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat
     4141 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     4201 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     4261 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     4321 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     4381 catggacgag ctgtacaagt aaagcggccg cgactctaga gtcgacctgc aggcatgcaa
     4441 gcttcaggta gccggctaac gttaacaacc ggtacctcta gaactatagc tagcatgcgc
     4501 aaatttaaag cgctgatatc gataaaataa aagattttat ttagtctcca gaaaaagggg
     4561 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     4621 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac
     4681 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     4741 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     4801 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     4861 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     4921 acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc
     4981 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     5041 tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg
     5101 gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact
     5161 gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag cgcagaaaga
     5221 gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5281 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5341 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5401 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5461 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5521 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5581 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5641 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5701 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5761 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5821 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5881 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5941 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6001 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6061 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6121 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6181 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6241 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6301 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6361 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6421 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6481 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6541 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6601 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6661 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6721 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6781 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6841 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6901 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6961 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7021 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7081 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7141 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7201 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7261 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7321 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     7381 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     7441 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     7501 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     7561 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     7621 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     7681 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     7741 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     7801 cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg agatggcgcc
     7861 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     7921 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
//
