LOCUS       sequence-358092-        8057 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 8057)
  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 8057)
  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-HA tagged-Mouse Mdm2 wt-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             98..614
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    678..1019
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1086..1502
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1432..1454
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1470..1486
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1531..1557
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3116..3665
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3260..3277)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3487..3506
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3668..3670,3671..3673,3674..4387)
                     /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(3668..3670,3671..3673,3674..4384)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3713..3734)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3974..3993)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4321..4342
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4549..5063
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5225..5241
                     /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(5249..5255,5256..5273,5274..5279))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5294..5315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5432..5449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5603..6191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5683..5702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6362..7153,7154..7222))
                     /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     6985..7004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7223..7327)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7395..7413
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7451..7473)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7573..7592
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7786..7808
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7936..7955
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa
       61 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
      121 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat
      181 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata
      241 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
      301 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
      361 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
      421 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
      481 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc
      541 cgaatcgtgg actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc
      601 gggggtcttt catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga
      661 cccccccgcc gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc
      721 gtgtttgtgc cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct
      781 gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga
      841 gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt
      901 ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc
      961 cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc
     1021 agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa
     1081 attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg
     1141 agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct
     1201 ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc
     1261 tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg
     1321 tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct
     1381 ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac
     1441 ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg
     1501 ccggaattcg attgggatcc agccatgggt tacccatacg acgtcccaga ctacgctacc
     1561 tgcaatacca acatgtctgt gtctaccgag ggtgctgcaa gcacctcaca gattccagct
     1621 tcggaacaag agactctggt tagaccaaaa ccattgcttt tgaagttgtt aaagtccgtt
     1681 ggagcgcaaa acgacactta cactatgaaa gagattatat tttatattgg ccagtatatt
     1741 atgactaaga ggttatatga cgagaagcag cagcacattg tgtattgttc aaatgatctc
     1801 ctaggagatg tgtttggagt cccgagtttc tctgtgaagg agcacaggaa aatatatgca
     1861 atgatctaca gaaatttagt ggctgtaagt cagcaagact ctggcacatc gctgagtgag
     1921 agcagacgtc agcctgaagg tgggagtgat ctgaaggatc ctttgcaagc gccaccagaa
     1981 gagaaacctt catcttctga tttaatttct agactgtcta cctcatctag aaggagatcc
     2041 attagtgaga cagaagagaa cacagatgag ctacctgggg agcggcaccg gaagcgccgc
     2101 aggtccctgt cctttgatcc gagcctgggt ctgtgtgagc tgagggagat gtgcagcggc
     2161 ggcagcagca gcagtagcag cagcagcagc gagtccacag agacgccctc gcatcaggat
     2221 cttgacgatg gcgtaagtga gcattctggt gattgcctgg atcaggattc agtttctgat
     2281 cagtttagcg tggaatttga agttgagtct ctggactcgg aagattacag cctgagtgac
     2341 gaagggcacg agctctcaga tgaggatgat gaggtctatc gggtcacagt ctatcagaca
     2401 ggagaaagcg atacagactc ttttgaaggg gatcctgaga tttccttagc tgactattgg
     2461 aagtgtacct catgcaatga aatgaatcct ccccttccat cacactgcaa aagatgctgg
     2521 acccttcgtg agaactggct tccagacgat aaggggaaag ataaagtgga aatctctgaa
     2581 aaagccaaac tggaaaactc agctcaggca gaagaaggct tggatgtgcc tgatggcaaa
     2641 aagctgacag agaatgatgc taaagagcca tgtgctgagg aggacagcga ggagaaggcc
     2701 gaacagacgc ccctgtccca ggagagtgac gactattccc aaccatcgac ttccagcagc
     2761 attgtttata gcagccaaga aagcgtgaaa gagttgaagg aggaaacgca ggacaaagac
     2821 gagagtgtgg aatctagctt ctccctgaat gccatcgaac catgtgtgat ctgccagggg
     2881 cggcctaaaa atggctgcat tgttcacggc aagactggac acctcatgtc atgtttcacg
     2941 tgtgcaaaga agctaaaaaa aagaaacaag ccctgcccag tgtgcagaca gccaatccaa
     3001 atgattgtgc taacttactt caactagctg acctgcttac aaaaatagaa ttttatatcg
     3061 ataatcacta gtgaattcgt taacctcgag cgggatcaat tccgcccccc ccctaacgtt
     3121 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     3181 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     3241 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     3301 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     3361 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     3421 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     3481 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     3541 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     3601 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     3661 taataccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     3721 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     3781 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     3841 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     3901 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     3961 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     4021 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     4081 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     4141 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     4201 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     4261 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     4321 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     4381 caagtaaagc ggccgcgact ctagagtcga cctgcaggca tgcaagcttc aggtagccgg
     4441 ctaacgttaa caaccggtac ctctagaact atagctagca tgcgcaaatt taaagcgctg
     4501 atatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     4561 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat
     4621 aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc
     4681 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     4741 ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     4801 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     4861 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg
     4921 cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc
     4981 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     5041 acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg
     5101 agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat
     5161 actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg ggagaattgt
     5221 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5281 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5341 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5401 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5461 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5521 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5581 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5641 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5701 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5761 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5821 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5881 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5941 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6001 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6061 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6121 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6181 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6241 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6301 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6361 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6421 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6481 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6541 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6601 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6661 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6721 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6781 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6841 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6901 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6961 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7021 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7081 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7141 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7201 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7261 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7321 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     7381 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     7441 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     7501 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     7561 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     7621 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     7681 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     7741 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     7801 gtaaaacgac ggcgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     7861 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     7921 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     7981 ggtgatgccg gccacgatgc gtccggcgta gaggcgatta gtccaatttg ttaaagacag
     8041 gatatcagtg gtccagg
//
