LOCUS       Exported                6455 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses human wt p53 and EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pIRES2-EGFP-p53 WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6455)
  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 6455)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6455
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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    1814..2400
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1981..1998)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2208..2227
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2401..3120
                     /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             2401..3117
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2446..2467)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2707..2726)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3054..3075
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3243..3364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3280..3299)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3334..3353
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3371..3826)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3508..3529)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3720..3739
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3853..3957
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3959..4316
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3982..4002)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4167..4302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4229..4248
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4351..5145
                     /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(4405..4424)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5015..5034
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5333..5352)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5377..5424
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5753..6341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6242..6261
                     /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 ccaagctgat ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg
     1861 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     1921 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     1981 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     2041 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     2101 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     2161 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     2221 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     2281 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     2341 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     2401 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2461 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2521 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2581 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     2641 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2701 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2761 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2821 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     2881 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2941 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3001 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3061 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     3121 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3181 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3241 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3301 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3361 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3421 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3481 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3541 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3601 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3661 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3721 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3781 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     3841 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3901 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     3961 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4021 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4081 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4141 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4201 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4261 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4321 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4381 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4441 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4501 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     4561 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4621 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4681 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4741 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4801 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4861 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4921 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4981 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5041 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5101 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5161 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5221 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5281 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5341 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5401 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5461 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5521 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     5581 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     5641 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5701 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5761 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5821 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5881 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5941 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6001 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6061 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6121 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6181 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6241 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6301 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6361 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6421 ctgattctgt ggataaccgt attaccgcca tgcat
//
