LOCUS       sequence-471593-        6450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human wt p53 and EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6450)
  AUTHORS   Lin SC, Karoly ED, Taatjes DJ
  TITLE     The human DeltaNp53 isoform triggers metabolic and gene expression 
            changes that activate mTOR and alter mitochondrial function.
  JOURNAL   Aging Cell. 2013 Oct;12(5):863-72. doi: 10.1111/acel.12108. Epub 
            2013 Jun 25.
  PUBMED    23734707
REFERENCE   2  (bases 1 to 6450)
  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"; doi: 
            "10.1111/acel.12108"; journalName: "Aging Cell"; date: "2013-10"; 
            volume: "12"; issue: "5"; pages: "863-72"
COMMENT     Sequence Label: pIRES2-EGFP-p53 WT
FEATURES             Location/Qualifiers
     source          1..6450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             613..1797
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /product="human tumor suppressor protein that is often 
                     mutated in cancers"
                     /label=p53
                     /note="responds to cellular stresses to regulate the 
                     expression of target genes"
                     /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML
                     SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSDV"
     misc_feature    join(1809..2383,2384..2386,2387..2395)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1976..1993)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2203..2222
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2396..2398,2399..2401,2402..3115)
                     /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(2396..2398,2399..2401,2402..3112)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2441..2462)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2702..2721)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3049..3070
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3238..3359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3275..3294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3329..3348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3366..3821)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3503..3524)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3715..3734
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3848..3952
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3954..4311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3954..4145
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3977..3997)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4162..4297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4346..5140
                     /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             4346..5140
                     /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(4400..4419)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5010..5029
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5328..5347)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5372..5419
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5748..6336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6237..6256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atatggagga gccgcagtca gatcctagcg tcgagccccc tctgagtcag
      661 gaaacatttt cagacctatg gaaactactt cctgaaaaca acgttctgtc ccccttgccg
      721 tcccaagcaa tggatgattt gatgctgtcc ccggacgata ttgaacaatg gttcactgaa
      781 gacccaggtc cagatgaagc tcccagaatg ccagaggctg ctccccccgt ggcccctgca
      841 ccagcagctc ctacaccggc ggcccctgca ccagccccct cctggcccct gtcatcttct
      901 gtcccttccc agaaaaccta ccagggcagc tacggtttcc gtctgggctt cttgcattct
      961 gggacagcca agtctgtgac ttgcacgtac tcccctgccc tcaacaagat gttttgccaa
     1021 ctggccaaga cctgccctgt gcagctgtgg gttgattcca cacccccgcc cggcacccgc
     1081 gtccgcgcca tggccatcta caagcagtca cagcacatga cggaggttgt gaggcgctgc
     1141 ccccaccatg agcgctgctc agatagcgat ggtctggccc ctcctcagca tcttatccga
     1201 gtggaaggaa atttgcgtgt ggagtatttg gatgacagaa acacttttcg acatagtgtg
     1261 gtggtgccct atgagccgcc tgaggttggc tctgactgta ccaccatcca ctacaactac
     1321 atgtgtaaca gttcctgcat gggcggcatg aaccggaggc ccatcctcac catcatcaca
     1381 ctggaagact ccagtggtaa tctactggga cggaacagct ttgaggtgcg tgtttgtgcc
     1441 tgtcctggga gagaccggcg cacagaggaa gagaatctcc gcaagaaagg ggagcctcac
     1501 cacgagctgc ccccagggag cactaagcga gcactgccca acaacaccag ctcctctccc
     1561 cagccaaaga agaaaccact ggatggagaa tatttcaccc ttcagatccg tgggcgtgag
     1621 cgcttcgaga tgttccgaga gctgaatgag gccttggaac tcaaggatgc ccaggctggg
     1681 aaggagccag gggggagcag ggctcactcc agccacctga agtccaaaaa gggtcagtct
     1741 acctcccgcc ataaaaaact catgttcaag acagaagggc ccgactcaga cgtctaggta
     1801 ccaagccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     1861 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     1921 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     1981 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     2041 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     2101 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     2161 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     2221 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     2281 cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     2341 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt
     2401 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2461 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2521 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2581 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2641 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2701 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2761 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2821 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2881 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2941 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3001 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3061 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     3121 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3181 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3241 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3301 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3361 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3421 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3481 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3541 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3601 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3661 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3721 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3781 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3841 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3961 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4021 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4081 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4141 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4201 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4261 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4321 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4381 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4441 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4501 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4561 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4621 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4681 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4741 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4801 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4861 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4921 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4981 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5041 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5101 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5161 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5221 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5281 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5341 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5401 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5461 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5521 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5581 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5641 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5701 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5761 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5821 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5881 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5941 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6001 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6061 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6121 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6181 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6241 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6301 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6361 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6421 tctgtggata accgtattac cgccatgcat
//
