LOCUS       sequence-358094-        6824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Role of Mdm2 variants in tumorigenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6824)
  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 6824)
  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 mutant M3 (399-489)-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..6824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             83..599
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    663..1004
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1071..1487
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1417..1439
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1455..1471
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1869..2418
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2013..2030)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2240..2259
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2421..2423,2424..2426,2427..3140)
                     /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(2421..2423,2424..2426,2427..3137)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2466..2487)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2727..2746)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3074..3095
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3302..3816
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3978..3994
                     /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(4002..4008,4009..4026,4027..4032))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4047..4068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4185..4202)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4356..4943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4436..4455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5114..5905,5906..5974))
                     /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     5737..5756
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5975..6079)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6147..6165
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6203..6225)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6325..6344
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6538..6560
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6688..6707
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa aagattttat
       61 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
      121 aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc
      181 aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt
      241 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag
      301 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      361 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      421 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc
      481 gcccgggtac ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggactcg
      541 ctgatccttg ggagggtctc ctcagattga ttgactgccc acctcggggg tctttcattt
      601 ggaggttcca ccgagatttg gagacccctg cccagggacc accgaccccc ccgccgggag
      661 gtaagctggc cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt tgtgccggca
      721 tctaatgttt gcgcctgcgt ctgtactagt tagctaacta gctctgtatc tggcggaccc
      781 gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact
      841 ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt
      901 caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat
      961 ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg
     1021 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg
     1081 ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca
     1141 accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa
     1201 cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta
     1261 agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga
     1321 cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc
     1381 ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc
     1441 cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga attcggatcc
     1501 accatgtcca gcagcattgt ttatagcagc caagaaagcg tgaaagagtt gaaggaggaa
     1561 acgcaggaca aagacgagag tgtggaatct agcttctccc tgaatgccat cgaaccatgt
     1621 gtgatctgcc aggggcggcc taaaaatggc tgcattgttc acggcaagac tggacacctc
     1681 atgtcatgtt tcacgtgtgc aaagaagcta aaaaaaagaa acaagccctg cccagtgtgc
     1741 agacagccaa tccaaatgat tgtgctaact tacttcaact agctgacctg ctcacaaaaa
     1801 tagaatttta tatcgaatca ctagtgaatt cgttaacctc gagcgggatc aattccgccc
     1861 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     1921 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     1981 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     2041 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     2101 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     2161 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     2221 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     2281 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     2341 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     2401 ttgaaaaaca cgataatacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2461 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2521 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2581 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     2641 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2701 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2761 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2821 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2881 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2941 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3001 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     3061 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3121 tggacgagct gtacaagtaa agcggccgcg actctagagt cgacctgcag gcatgcaagc
     3181 ttcaggtagc cggctaacgt taacaaccgg tacctctaga actatagcta gcatgcgcaa
     3241 atttaaagcg ctgatatcga taaaataaaa gattttattt agtctccaga aaaagggggg
     3301 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     3361 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
     3421 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     3481 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
     3541 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     3601 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     3661 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa
     3721 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     3781 tgagtgattg actacccgtc agcgggggtc tttcatgggt aacagtttct tgaagttgga
     3841 gaacaacatt ctgagggtag gagtcgaata ttaagtaatc ctgactcaat tagccactgt
     3901 tttgaatcca catactccaa tactcctgaa atagttcatt atggacagcg cagaaagagc
     3961 tggggagaat tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4021 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4081 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4141 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4201 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4261 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4321 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4381 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4441 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4501 accggatacc tgtccgcctt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4561 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4621 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4681 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4741 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4801 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4861 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4921 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4981 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5041 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5101 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5161 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5221 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5281 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5341 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5401 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5461 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5521 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5581 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5641 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5701 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5761 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5821 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5881 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5941 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6001 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6061 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     6121 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     6181 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     6241 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     6301 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     6361 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     6421 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     6481 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     6541 agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa
     6601 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     6661 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     6721 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt
     6781 tgttaaagac aggatatcag tggtccaggc tctagttttg actc
//
