LOCUS       sequence-358090-        7124 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 7124)
  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 7124)
  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 M2 (299-489)-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..7124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    189..530
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             597..1013
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     943..965
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     981..997
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1694..2243
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1838..1855)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2065..2084
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2246..2248,2249..2251,2252..2965)
                     /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(2246..2248,2249..2251,2252..2962)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2291..2312)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2552..2571)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2899..2920
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3127..3641
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3803..3819
                     /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(3827..3833,3834..3851,3852..3857))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3872..3893
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4010..4027)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4181..4769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4261..4280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4940..5731,5732..5800))
                     /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     5563..5582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5801..5905)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5973..5991
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6029..6051)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6151..6170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6364..6386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6514..6533
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(6733..7124,1..125)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgcgtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgaatcgtg
       61 gactcgctga tccttgggag ggtctcctca gattgattga ctgcccacct cgggggtctt
      121 tcatttggag gttccaccga gatttggaga cccctgccca gggaccaccg acccccccgc
      181 cgggaggtaa gctggccagc ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg
      241 ccggcatcta atgtttgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc
      301 ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca
      361 gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga
      421 ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
      481 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
      541 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
      601 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
      661 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
      721 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
      781 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
      841 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
      901 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
      961 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggaattc
     1021 ggatccacca tggactattg gaagtgtacc tcatgcaatg aaatgaatcc tccccttcca
     1081 tcacactgca aaagatgctg gacccttcgt gagaactggc ttccagacga taaggggaaa
     1141 gataaagtgg aaatctctga aaaagccaaa ctggaaaact cagctcaggc agaagaaggc
     1201 ttggatgtgc ctgatggcaa aaagctgaca gagaatgatg ctaaagagcc atgtgctgag
     1261 gaggacagcg aggagaaggc cgaacagacg cccctgtccc aggagagtga cgactattcc
     1321 caaccatcga cttccagcag cattgtttat agcagccaag aaagcgtgaa agagttgaag
     1381 gaggaaacgc aggacaaaga cgagagtgtg gaatctagct tctccctgaa tgccatcgaa
     1441 ccatgtgtga tctgccaggg gcggcctaaa aatggctgca ttgttcacgg caagactgga
     1501 cacctcatgt catgtttcac gtgtgcaaag aagctaaaaa aaagaaacaa gccctgccca
     1561 gtgtgcagac agccaatcca aatgattgtg ctaacttact tcaactagct gacctgctca
     1621 caaaaataga attttatatc gatcactagt gaattcgtta acctcgagcg ggatcaattc
     1681 cgcccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     1741 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     1801 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     1861 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     1921 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     1981 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     2041 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     2101 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     2161 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     2221 ttcctttgaa aaacacgata ataccatggt gagcaagggc gaggagctgt tcaccggggt
     2281 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     2341 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     2401 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     2461 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2521 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2581 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2641 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     2701 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     2761 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2821 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     2881 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2941 cggcatggac gagctgtaca agtaaagcgg ccgcgactct agagtcgacc tgcaggcatg
     3001 caagcttcag gtagccggct aacgttaaca accggtacct ctagaactat agctagcatg
     3061 cgcaaattta aagcgctgat atcgataaaa taaaagattt tatttagtct ccagaaaaag
     3121 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     3181 aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga
     3241 gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     3301 gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca
     3361 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     3421 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     3481 acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta
     3541 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     3601 tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag
     3661 ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc
     3721 actgttttga atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa
     3781 agagctgggg agaattgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3841 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3901 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3961 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4021 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4081 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4141 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4201 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4261 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4321 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4381 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4441 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4501 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4561 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4621 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4681 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4741 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4801 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4861 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4921 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4981 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5041 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5101 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5161 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5221 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5281 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5341 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5401 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5461 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5521 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5581 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5641 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5701 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5761 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5821 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5881 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5941 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     6001 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     6061 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     6121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     6181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     6241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     6301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     6361 tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc
     6421 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     6481 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     6541 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt
     6601 ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca
     6661 ccagctgaag cctatagagt acgagccata gataaaataa aagattttat ttagtctcca
     6721 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     6781 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
     6841 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     6901 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
     6961 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     7021 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     7081 agagcccaca acccctcact cggcgcgcca gtcctccgat agac
//
