LOCUS       sequence-72827-e        7588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human p53 deltaN linked to full length p53 with C terminal
            His tag and also expresses EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7588)
  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 7588)
  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-deltaNp53:WTp53
FEATURES             Location/Qualifiers
     source          1..7588
                     /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             1689..1709
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1719..2897
                     /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
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     CDS             2901..2924
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     misc_feature    join(2947..3521,3522..3524,3525..3533)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3114..3131)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3341..3360
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3534..3536,3537..3539,3540..4253)
                     /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(3534..3536,3537..3539,3540..4250)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3579..3600)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3840..3859)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4187..4208
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4376..4497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4413..4432)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4467..4486
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4504..4959)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4641..4662)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4853..4872
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4986..5090
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5092..5449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5092..5283
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5115..5135)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5300..5435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5484..6278
                     /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             5484..6278
                     /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(5538..5557)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6148..6167
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6466..6485)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6510..6557
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6886..7474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7375..7394
                     /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 tatggatgat gatttgatgc tgtccccgga cgatattgaa caatggttca
      661 ctgaagaccc aggtccagat gaagctccca gaatgccaga ggctgctccc cccgtggccc
      721 ctgcaccagc agctcctaca ccggcggccc ctgcaccagc cccctcctgg cccctgtcat
      781 cttctgtccc ttcccagaaa acctaccagg gcagctacgg tttccgtctg ggcttcttgc
      841 attctgggac agccaagtct gtgacttgca cgtactcccc tgccctcaac aagatgtttt
      901 gccaactggc caagacctgc cctgtgcagc tgtgggttga ttccacaccc ccgcccggca
      961 cccgcgtccg cgccatggcc atctacaagc agtcacagca catgacggag gttgtgaggc
     1021 gctgccccca ccatgagcgc tgctcagata gcgatggtct ggcccctcct cagcatctta
     1081 tccgagtgga aggaaatttg cgtgtggagt atttggatga cagaaacact tttcgacata
     1141 gtgtggtggt gccctatgag ccgcctgagg ttggctctga ctgtaccacc atccactaca
     1201 actacatgtg taacagttcc tgcatgggcg gcatgaaccg gaggcccatc ctcaccatca
     1261 tcacactgga agactccagt ggtaatctac tgggacggaa cagctttgag gtgcgtgttt
     1321 gtgcctgtcc tgggagagac cggcgcacag aggaagagaa tctccgcaag aaaggggagc
     1381 ctcaccacga gctgccccca gggagcacta agcgagcact gcccaacaac accagctcct
     1441 ctccccagcc aaagaagaaa ccactggatg gagaatattt cacccttcag atccgtgggc
     1501 gtgagcgctt cgagatgttc cgagagctga atgaggcctt ggaactcaag gatgcccagg
     1561 ctgggaagga gccagggggg agcagggctc actccagcca cctgaagtcc aaaaagggtc
     1621 agtctacctc ccgccataaa aaactcatgt tcaagacaga agggccagac tctgacggtg
     1681 gcggagggga aaatctgtat ttccagggag gcggaggtat ggaagagccg cagtcagatc
     1741 ctagcgtcga gccccctctg agtcaggaaa cattttcaga cctatggaaa ctacttcctg
     1801 aaaacaacgt tctgtccccc ttgccgtccc aagcaatgga tgatttgatg ctgtccccgg
     1861 acgatattga acaatggttc actgaagacc caggtccaga tgaagctccc agaatgccag
     1921 aggctgctcc ccccgtggcc cctgcaccag cagctcctac accggcggcc cctgcaccag
     1981 ccccctcctg gcccctgtca tcttctgtcc cttcccagaa aacctaccag ggcagctacg
     2041 gtttccgtct gggcttcttg cattctggga cagccaagtc tgtgacttgc acgtactccc
     2101 ctgccctcaa caagatgttt tgccaactgg ccaagacctg ccctgtgcag ctgtgggttg
     2161 attccacacc cccgcccggc acccgcgtcc gcgccatggc catctacaag cagtcacagc
     2221 acatgacgga ggttgtgagg cgctgccccc accatgagcg ctgctcagat agcgatggtc
     2281 tggcccctcc tcagcatctt atccgagtgg aaggaaattt gcgtgtggag tatttggatg
     2341 acagaaacac ttttcgacat agtgtggtgg tgccctatga gccgcctgag gttggctctg
     2401 actgtaccac catccactac aactacatgt gtaacagttc ctgcatgggc ggcatgaacc
     2461 ggaggcccat cctcaccatc atcacactgg aagactccag tggtaatcta ctgggacgga
     2521 acagctttga ggtgcgtgtt tgtgcctgtc ctgggagaga ccggcgcaca gaggaagaga
     2581 atctccgcaa gaaaggggag cctcaccacg agctgccccc agggagcact aagcgagcac
     2641 tgcccaacaa caccagctcc tctccccagc caaagaagaa accactggat ggagaatatt
     2701 tcacccttca gatccgtggg cgtgagcgct tcgagatgtt ccgagagctg aatgaggcct
     2761 tggaactcaa ggatgcccag gctgggaagg agccaggggg gagcagggct cactccagcc
     2821 acctgaagtc caaaaagggt cagtctacct cccgccataa aaaactcatg ttcaagacag
     2881 aagggcccga ctcagacgtg catcaccatc accatcacca tcacgtctag gtaccaagct
     2941 gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     3001 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     3061 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     3121 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     3181 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     3241 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     3301 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     3361 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     3421 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     3481 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtga
     3541 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     3601 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     3661 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     3721 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     3781 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     3841 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     3901 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     3961 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     4021 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     4081 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     4141 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     4201 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     4261 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     4321 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     4381 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4441 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     4501 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     4561 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     4621 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4681 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4741 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4801 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4861 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4921 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     4981 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5041 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     5101 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     5161 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     5221 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     5281 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5341 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     5401 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     5461 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5521 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5581 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5641 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     5701 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5761 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5821 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5881 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5941 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     6001 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     6061 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6121 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6181 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6241 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     6301 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6361 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6421 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     6481 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     6541 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     6601 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     6661 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     6721 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6781 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6841 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6901 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6961 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7021 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7081 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7141 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7201 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7261 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7321 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     7381 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7441 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7501 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     7561 tgtggataac cgtattaccg ccatgcat
//
