LOCUS       sequence-307568-        9151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP tagged Mdm2 from MDM2 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9151)
  AUTHORS   Lahav G, Rosenfeld N, Sigal A, Geva-Zatorsky N, Levine AJ, Elowitz 
            MB, Alon U
  TITLE     Dynamics of the p53-Mdm2 feedback loop in individual cells.
  JOURNAL   Nat Genet. 2004 Feb;36(2):147-50. Epub 2004 Jan 18.
  PUBMED    14730303
REFERENCE   2  (bases 1 to 9151)
  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: "Nat 
            Genet."; date: "2004-02"; volume: "36(2)"; pages: "147-50. Epub 2004
            Jan 18"
COMMENT     Sequence Label: MDM2p-Mdm2-YFP
FEATURES             Location/Qualifiers
     source          1..9151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      5056..5065
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5062..5064,5065..5067,5068..5781)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5107..5128)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5368..5387)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5715..5736
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5904..6025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5941..5960)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5995..6014
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6032..6487)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6169..6190)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6381..6400
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6514..6618
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6620..6977
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6620..6811
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6643..6663)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6828..6963
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7012..7806
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             7012..7806
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7066..7085)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7676..7695
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7994..8013)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8038..8085
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8414..9002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8903..8922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actaccgaat tctgttctgt gacccataac ttaaaactaa cccaactttt cattccctcc
       61 agactttttt tggtaagcag tatcatcatg ctttaggtgg acactgagtc atactgctgt
      121 ggtagtgctg gtacacgtag ttttgaatac caggttcttg agtgaatcct ttttacttct
      181 ctctctctcc cccacccccg cttctccctt ctgccttcat ttctgaccct tcttttctgg
      241 ttccattaga aaaggactaa tcttatcagt tctgcctttt tttttctgct ctaataattt
      301 tgtcaagaga attttttcaa agcacaaggc caatgagaag ggggaaagtc ggtccagaag
      361 gcagccggcc cgactattct gagtttgaat ctactcttgg tggtcctgtt aaaatacagt
      421 ttgaagcaaa tcacttgttt ttatgcttct aatgtctact ctgtcgaaag cccaaatttc
      481 cttgcttggc atttaaggag actccaagtt cttaaaactt cttgactttc atgcaagtca
      541 aacggattta tttattgtat ctcaaacagt cctcttatat tttcctttct gtattttttg
      601 ataattttct tgtgcctgga atattctctg cttttattta agtctcacca tctttcaaga
      661 cacaaaatat gtgtgatctc tgaggccttt gtgaactgcc tccagcattt actaagcaat
      721 taattctgtc tctgaacttt tatagcacaa acccattcgt tcacttgata tttaacaaat
      781 atcaacctct atcacttatc tctttttttt tttttttttc ttctgagatg gagtcttgct
      841 ctgtcaccca ggctggactg cagtggcgcg atctccgctc actgcaacct ctgcctcccg
      901 agatcaagcg attcttctgc ctcagcctcc caagtggcta ggattacagg cgcccgccac
      961 ggggcccggc taatttttgt atttttagta gagacaaggt ttcaccatct tggtcaggct
     1021 ggtctcgaac tcctaacctc gtgatccgcc cacctcagcc tcccaaagtg ctgggattac
     1081 aagcgtgagc caccgtgcgc agcctatcac ttatcttttc atgcttacat tttctcactg
     1141 cttgagattt tctttcatcc agggtcaagc actgagtcta ttagaaaccc cactgtatat
     1201 gatctgatga agatgagatt atctttttaa aagccacata ttaaatctct taagttaata
     1261 atattggcaa tcaacttaaa tcaactgagt gaccactatg tttaaggaag tttcctttct
     1321 ggtaggctgg gaagcgggag aagggaagga tataacttta taaaaaaaaa aagctgcaga
     1381 agggaaggat ataactttat aaaaaaaggt ccttgttcac aaggagctaa tcacttactg
     1441 gccagagaag aaacgtgctc cttctttgca atggctgtgt ttttgggttt agctaactca
     1501 attctttatg tcctcaattt atgagaccgc atcaaaaacg tttttgccac atctctgtgg
     1561 gaaagtaggt gagtcagaat gagtgcataa tatataagta tatattttaa tgataatgaa
     1621 tgtgtagatt aacaaaccca aactttgttt ttgttttttt gagagtcttg ccctgtcgcc
     1681 caggctggag tgcaatggca tgatcttggc tcactgcagc ctctgcctcc cgggttcaca
     1741 ggattgtcct gcctcagcct ccggagtaga gtagctggga ttacaggtga ccgccaccac
     1801 gcccgactaa tttttgtagt ttttagtaga gacggcgttt cccatgttgg ccaggttggt
     1861 cttgaactcc tgacctcaag tgatccaccc gcctcggcct acccaaagtg atgggattac
     1921 aagcgtgagc caccgcgccc agccaaaccc aaacattcta aaagcgcaga gtaaccgctc
     1981 ccctccccgc caacggtaac cagcatcttc gttctccaac cagaacagct gtatgcaccg
     2041 gtgtataagc ctgccttaag tgctattttt aatctttttt aaaaacaggc cttaaaataa
     2101 ctcatttggg tacaactcca gcggctggtg gagtgggctt aatatattaa acagctgtta
     2161 attttggttt ctttttggta acagcgacac ggagatccgc atttgtcgca gtttccaccg
     2221 cgggcgggaa gtgtaaacac aagaaataca aacatagcgc aacggctaaa ggagtgtcac
     2281 agcgccaaac cttgctggct ccgcgccagg ccgagcccca ccctctcagc tcgcggccaa
     2341 cacccccacc ccgcctcaca gcccgccgcg cccgcggggc gacacccccc actccatcat
     2401 cccggaggtg gtgcggccga gccccggacc caattggcgg aagcggggcc ggttgtgtgc
     2461 gcgcgcacaa atgcccggat gcgccgcgac gacccgcgcg ctcccctcgg gcggtagggg
     2521 gcgcgcaccg aggcaccgcg gcgagcttgg ctgcttctgg ggcctgtgtg gccctgtgtg
     2581 tcggaaagat ggagcaagaa gccgagcccg aggggcggcc gcgacccctc tgaccgagat
     2641 cctgctgctt tcgcagccag gagcaccgtc cctccccgga ttagtgcgta cgagcgccca
     2701 gtgccctggc ccggagagtg gaatgatccc cgaggcccag ggcgtcgtgc ttccgcgcgc
     2761 cccgtgaagg aaactgggga gtcttgaggg acccccgact ccaagcgcga aaaccccgga
     2821 tggtgaggag caggtactgg cccggcagcg agcggtcact tttgggtctg ggctctgacg
     2881 gtgtcccctc tatcgctggt tcccagcctc tgcccgttcg cagcctttgt gcggttcgtg
     2941 gctgggggct cggggcgcgg ggcgcggggc atggggcacg tggctttgcg gaggttttgt
     3001 tggactgggg ctaggcagtc gccgccaggg aggagggcgg gatttcggac ggctctcgcg
     3061 gcggtggggg tgggggtggt tcggaggtct ccgcgggagt tcagggtaaa ggtcacgggg
     3121 gccgggggct gcggggccgc tgcggcgcgg gaggtccgga tgatcgcagg tgcctgtcgg
     3181 gtcactagtg tgaacgctgc gcgtagtctg ggcgggattg ggccggttca gtgggcaggt
     3241 tgactcagct tttcctcttg agctggtcaa gttcagacac gttccgaaac tgcagtaaaa
     3301 ggagttaagt cctgacttgt ctccagctgg ggctatttaa accatgcatt ttcccagctg
     3361 tgttcagtgg cgattggagg gtagacctgt gggcacggac gcacgccact ttttctctgc
     3421 tgatccaggt aagcaccgac ttgcttgtag ctttagtttt aactgttgtt tatgttcttt
     3481 atatatgatg tattttccac agatgtttca tgatttccag ttttcatcgt gtcttttttt
     3541 tccttgtagg caaatgtgct cgagcaaatg tgcaatacca acatgtctgt acctactgat
     3601 ggtgctgtaa ccacctcaca gattccagct tcggaacaag agaccctggt tagaccaaag
     3661 ccattgcttt tgaagttatt aaagtctgtt ggtgcacaaa aagacactta tactatgaaa
     3721 gaggttcttt tttatcttgg ccagtatatt atgactaaac gattatatga tgagaagcaa
     3781 caacatattg tatattgttc aaatgatctt ctaggagatt tgtttggcgt gccaagcttc
     3841 tctgtgaaag agcacaggaa aatatatacc atgatctaca ggaacttggt agtagtcaat
     3901 cagcaggaat catcggactc aggtacatct gtgagtgaga acaggtgtca ccttgaaggt
     3961 gggagtgatc aaaaggacct tgtacaagag cttcaggaag agaaaccttc atcttcacat
     4021 ttggtttcta gaccatctac ctcatctaga aggagagcaa ttagtgagac agaagaaaat
     4081 tcagatgaat tatctggtga acgacaaaga aaacgccaca aatctgatag tatttccctt
     4141 tcctttgatg aaagcctggc tctgtgtgta ataagggaga tatgttgtga aagaagcagt
     4201 agcagtgaat ctacagggac gccatcgaat ccggatcttg atgctggtgt aagtgaacat
     4261 tcaggtgatt ggttggatca ggattcagtt tcagatcagt ttagtgtaga atttgaagtt
     4321 gaatctctcg actcagaaga ttatagcctt agtgaagaag gacaagaact ctcagatgaa
     4381 gatgatgagg tatatcaagt tactgtgtat caggcagggg agagtgatac agattcattt
     4441 gaagaagatc ctgaaatttc cttagctgac tattggaaat gcacttcatg caatgaaatg
     4501 aatccccccc ttccatcaca ttgcaacaga tgttgggccc ttcgtgagaa ttggcttcct
     4561 gaagataaag ggaaagataa aggggaaatc tctgagaaag ccaaactgga aaactcaaca
     4621 caagctgaag agggctttga tgttcctgat tgtaaaaaaa ctatagtgaa tgattccaga
     4681 gagtcatgtg ttgaggaaaa tgatgataaa attacacaag cttcacaatc acaagaaagt
     4741 gaagactatt ctcagccatc aacttctagt agcattattt atagcagcca agaagatgtg
     4801 aaagagtttg aaagggaaga aacccaagac aaagaagaga gtgtggaatc tagtttgccc
     4861 cttaatgcca ttgaaccttg tgtgatttgt caaggtcgac ctaaaaatgg ttgcattgtc
     4921 catggcaaaa caggacatct tatggcctgc tttacatgtg caaagaagct aaagaaaagg
     4981 aataagccct gcccagtatg tagacaacca attcaaatga ttgtgctaac ttatttcccc
     5041 cgggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     5101 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     5161 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     5221 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc
     5281 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     5341 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     5401 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     5461 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     5521 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     5581 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     5641 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     5701 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     5761 atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat
     5821 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5881 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5941 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6001 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     6061 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     6121 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     6181 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     6241 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     6301 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     6361 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6421 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6481 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6541 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6601 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     6661 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6721 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6781 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     6841 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6901 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6961 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     7021 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     7081 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     7141 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     7201 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     7261 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     7321 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     7381 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     7441 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     7501 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     7561 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     7621 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     7681 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     7741 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     7801 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     7861 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     7921 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     7981 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     8041 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     8101 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     8161 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     8221 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     8281 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     8341 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8401 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     8461 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     8521 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     8581 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8641 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8701 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     8761 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8821 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8881 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8941 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     9001 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     9061 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     9121 tattactagc atcaagctta tcgataccgt c
//
