LOCUS       sequence-358089-        8047 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 8047)
  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 8047)
  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 wild type-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     393..414
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    462..1011
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(621..640)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     850..867
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(2615..3031)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2631..2647)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2663..2685)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3098..3439
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             3503..4019
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4219..4238)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4366..4388)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4582..4601)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4701..4723
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4761..4779)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4847..4951
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4952..5020,5021..5812)
                     /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     complement(5170..5189)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5983..6571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6472..6491
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6725..6742
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6859..6880
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6895..6900,6901..6918,6919..6925)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6933..6949
                     /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)."
     LTR             7111..7625
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(7787..8047,1..453,454..456,457..459))
                     /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             complement(join(7790..8047,1..453,454..456,457..459))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7832..7853)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
       61 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
      121 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
      181 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
      241 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
      301 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
      361 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
      421 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtattatcgt gtttttcaaa
      481 ggaaaaccac gtccccgtgg ttcggggggc ctagacgttt ttttaacctc gactaaacac
      541 atgtaaagca tgtgcaccga ggccccagat cagatcccat acaatggggt accttctggg
      601 catccttcag ccccttgttg aatacgcttg aggagagcca tttgactctt tccacaacta
      661 tccaactcac aacgtggcac tggggttgtg ccgcctttgc aggtgtatct tatacacgtg
      721 gcttttggcc gcagaggcac ctgtcgccag gtggggggtt ccgctgcctg caaagggtcg
      781 ctacagacgt tgtttgtctt caagaagctt ccagaggaac tgcttccttc acgacattca
      841 acagaccttg cattcctttg gcgagagggg aaagacccct aggaatgctc gtcaagaaga
      901 cagggccagg tttccgggcc ctcacattgc caaaagacgg caatatggtg gaaaataaca
      961 tatagacaaa cgcacaccgg ccttattcca agcggcttcg gccagtaacg ttaggggggg
     1021 ggcggaattg atcccgctcg aggttaacga attcactagt gattatcgat ataaaattct
     1081 atttttgtga gcaggtcagc tagttgaagt aagttagcac aatcatttgg attggctgtc
     1141 tgcacactgg gcagggcttg tttctttttt ttagcttctt tgcacacgtg aaacatgaca
     1201 tgaggtgtcc agtcttgccg tgaacaatgc agccattttt aggccgcccc tggcagatca
     1261 cacatggttc gatggcattc agggagaagc tagattccac actctcgtct ttgtcctgcg
     1321 tttcctcctt caactctttc acgctttctt ggctgctata aacaatgctg ctggaagtcg
     1381 atggttggga atagtcgtca ctctcctggg acaggggcgt ctgttcggcc ttctcctcgc
     1441 tgtcctcctc agcacatggc tctttagcat cattctctgt cagctttttg ccatcaggca
     1501 catccaagcc ttcttctgcc tgagctgagt tttccagttt ggctttttca gagatttcca
     1561 ctttatcttt ccccttatcg tctggaagcc agttctcacg aagggtccag catcttttgc
     1621 agtgtgatgg aaggggagga ttcatttcat tgcatgaggt acacttccaa tagtcagcta
     1681 aggaaatctc aggatcccct tcaaaagagt ctgtatcgct ttctcctgtc tgatagactg
     1741 tgacccgata gacctcatca tcctcatctg agagctcgtg cccttcgtca ctcaggctgt
     1801 aatcttccga gtccagagac tcaacttcaa attccacgct aaactgatca gaaactgaat
     1861 cctgatccag gcaatcacca gaatgctcac ttacgccatc gtcaagatcc tgatgcgagg
     1921 gcgtctctgt ggactcgctg ctgctgctgc tactgctgct gctgccgccg ctgcacatct
     1981 ccctcagctc acacagaccc aggctcggat caaaggacag ggacctgcgg cgcttccggt
     2041 gccgctcccc aggtagctca tctgtgttct cttctgtctc actaatggat ctccttctag
     2101 atgaggtaga cagtctagaa attaaatcag aagatgaagg tttctcttct ggtggcgctt
     2161 gcaaaggatc cttcagatca ctcccacctt caggctgacg tctgctctca ctcagcgatg
     2221 tgccagagtc ttgctgactt acagccacta aatttctgta gatcattgca tatattttcc
     2281 tgtgctcctt cacagagaaa ctcgggactc caaacacatc tcctaggaga tcatttgaac
     2341 aatacacaat gtgctgctgc ttctcgtcat ataacctctt agtcataata tactggccaa
     2401 tataaaatat aatctctttc atagtgtaag tgtcgttttg cgctccaacg gactttaaca
     2461 acttcaaaag caatggtttt ggtctaacca gagtctcttg ttccgaagct ggaatctgtg
     2521 aggtgcttgc agcaccctcg gtagacacag acatgttggt attgcacatt ggcctggagg
     2581 ccgcaggaga agcgcggtga tcgaatcgaa ttccggcgcc tagagaagga gtgagggctg
     2641 gataaaggga ggatcgaggc ggggtcgaac gaggaggttc aagggggaga gacggggcgg
     2701 atggaggaag aggaggcgga ggcttagggt gtacaaaggg cttgacccag ggaggggggt
     2761 caaaagccaa ggcttcccag gtcacgatgt aggggacctg gtctgggtgt ccatgcgggc
     2821 caggtgaaaa gaccttgatc ttaacctggg tgatgaggtc tcggttaaag gtgccgtctc
     2881 gcggccatcc gacgttaaag gttggccatt ctgcagagca gaaggtaacc caacgtctct
     2941 tcttgacatc taccgactgg ttgtgagcga tccgctcgac atctttccag tgacctaagg
     3001 tcaaacttaa gggagtggta acagtctggc cctaattttc agacaaatac agaaacacag
     3061 tcagacagag acaacacaga acgatgctgc agcgctgcag cagacaagac gcgcggcttc
     3121 ggttccaaac cgaaagcaaa aattcagacg gaggcgggaa ctgttttagg ttctcgtctc
     3181 ctaccagaac cacatatcct gacggggtcg gattccacat cgactccctt cctcaggtcg
     3241 ggccacaaaa acggccccca aagtccctgg gacgtctccc agggttgcgg ccgggtgttc
     3301 agaactcgtc agttccacca cgggtccgcc agatacagag ctagttagct aactagtaca
     3361 gacgcaggcg caaacattag atgccggcac aaacacgcac aaagacagag acagacacga
     3421 aacgaccgct ggccagctta cctcccggcg ggggggtcgg tggtccctgg gcaggggtct
     3481 ccaaatctcg gtggaacctc caaatgaaag acccccgagg tgggcagtca atcaatctga
     3541 ggagaccctc ccaaggatca gcgagtccac gattcggatg caaacagcaa gaggctttat
     3601 tgggaatacg ggtacccggg cgacgcagtc tatcggagga ctggcgcgcc gagtgagggg
     3661 ttgtgggctc ttttattgag ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa
     3721 ctgattggtt agttcaaata aggcacaggg tcatttcagg tccttggggc accctggaaa
     3781 catctgatgg ttctctagaa actgctgagg gcgggaccgc atctggggac catctgttct
     3841 tggccctgag ccggggcagg aactgcttac cacagatatc ctgtttggcc catattctgc
     3901 tgtctctctg ttcctaacct tgatctgaac ttctctattc tcagttatgt attttccatg
     3961 ccttgcaaaa tggcgttact taagctagct tgccaaacct acaggtgggg tctttcattc
     4021 cccccttttt ctggagacta aataaaatct tttattttat ctatggctcg tactctatag
     4081 gcttcagctg gtgatattgt tgagtcaaaa ctagagcctg gaccactgat atcctgtctt
     4141 taacaaattg gactaatcgc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     4201 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     4261 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     4321 gactgttggg cgccatctcc ttgcatgcac cattccttgc gccgtcgttt tacaacgtcg
     4381 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     4441 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     4501 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4561 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4621 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4681 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4741 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4801 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4861 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4921 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4981 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5041 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5101 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5161 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5221 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5281 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5341 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5401 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5461 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5521 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5581 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5641 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5701 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5761 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5821 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5881 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5941 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6001 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6061 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6121 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6181 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6241 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6301 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6361 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6421 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6481 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6541 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6601 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6661 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6721 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     6781 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6841 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     6901 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacaattc
     6961 tccccagctc tttctgcgct gtccataatg aactatttca ggagtattgg agtatgtgga
     7021 ttcaaaacag tggctaattg agtcaggatt acttaatatt cgactcctac cctcagaatg
     7081 ttgttctcca acttcaagaa actgttaccc atgaaagacc cccgctgacg ggtagtcaat
     7141 cactcagagg agaccctccc aaggaacagc gagaccacaa gtcggatgca actgcaagag
     7201 ggtttattgg atacacgggt acccgggcga cgcagtctat cggaggactg gcgcgccgag
     7261 tgaggggttg tgggctcttt tattgagctc ggggagcaga agcgcgcgaa cagaagcgag
     7321 aagcgaactg attggttagt tcaaataagg cacagggtca tttcaggtcc ttggggcacc
     7381 ctggaaacat ctgatggttc tctagaaact gctgagggcg ggaccgcatc tggggaccat
     7441 ctgttcttgg ccctgagccg gggcaggaac tgcttaccac agatatcctg tttggcccat
     7501 attctgctgt ctctctgttc ctaaccttga tctgaacttc tctattctca gttatgtatt
     7561 ttccatgcct tgcaaaatgg cgttacttaa gctagcttgc caaacctaca ggtggggtct
     7621 ttcattcccc cctttttctg gagactaaat aaaatctttt attttatcga tatcagcgct
     7681 ttaaatttgc gcatgctagc tatagttcta gaggtaccgg ttgttaacgt tagccggcta
     7741 cctgaagctt gcatgcctgc aggtcgactc tagagtcgcg gccgctttac ttgtacagct
     7801 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     7861 gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg ttgtcgggca
     7921 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     7981 tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt
     8041 cggccat
//
