LOCUS       sequence-387840-        6801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of 3xHA tagged wild-type p53.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6801)
  AUTHORS   Raj N, Wang M, Seoane JA, Zhao RL, Kaiser AM, Moonie NA, Demeter J, 
            Boutelle AM, Kerr CH, Mulligan AS, Moffatt C, Zeng SX, Lu H, Barna 
            M, Curtis C, Chang HY, Jackson PK, Attardi LD
  TITLE     The Mettl3 epitranscriptomic writer amplifies p53 stress responses.
  JOURNAL   Mol Cell. 2022 Jul 7;82(13):2370-2384.e10. doi: 
            10.1016/j.molcel.2022.04.010. Epub 2022 May 4.
  PUBMED    35512709
REFERENCE   2  (bases 1 to 6801)
  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: 
            "10.1016/j.molcel.2022.04.010"; journalName: "Mol Cell"; date: 
            "2022-07-7- 7"; volume: "82"; issue: "13"; pages: "2370-2384.e10"
COMMENT     Sequence Label: pcDNA3.1-3XHA-p53
FEATURES             Location/Qualifiers
     source          1..6801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(15..35)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        20..349
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      200..335
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     262..281
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             416..1210
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(470..489)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1080..1099
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1386..1507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1423..1442)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1477..1496
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1556..1572)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1556..1572)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1569..1591)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1580..1596
                     /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(1604..1610,1611..1628,1629..1634))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1649..1670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1787..1804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1958..2546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2038..2057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2717..3508,3509..3577))
                     /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     3340..3359
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3578..3682)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3757..3776)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3948..4327
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4328..4531
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4482..4502
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4576..4595
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4576..4594
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4673..4682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4694..4720
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4724..4750
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4754..4780
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(5177..5188)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             6018..6047
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6063..6080
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6103..6120)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6109..6333
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6379..6801,1..6)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6466..6485)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6676..6697
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
       61 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
      121 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
      181 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
      241 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
      301 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
      361 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
      421 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
      481 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
      541 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
      601 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
      661 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
      721 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
      781 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
      841 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
      901 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
      961 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     1021 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     1081 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     1141 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     1201 gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc
     1261 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     1321 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     1381 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1441 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1501 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     1561 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1621 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1681 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1741 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     1801 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1861 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1921 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1981 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2041 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2101 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2161 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2221 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2281 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2341 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2401 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2461 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2521 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2581 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2641 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2701 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2761 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2821 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     2881 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2941 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3001 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3061 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3121 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3181 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3241 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3301 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3361 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3421 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3481 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3541 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3601 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3661 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     3721 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     3781 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     3841 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     3901 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
     3961 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     4021 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     4081 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     4141 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     4201 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     4261 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     4321 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     4381 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     4441 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     4501 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
     4561 ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct agttaagctt
     4621 ggtaccgagc tcggatccac tagtccagtg tggtggaatt ctgcagataa acgccaccat
     4681 ggggcaacaa ggatacccgt atgatgttcc ggattacgct ggctacccat acgacgtccc
     4741 agactacgct ggctacccat acgacgtccc agactacgct caaggagtgc ggccgcgggg
     4801 gcgcgccact gccatggagg agtcacagtc ggatatcagc ctcgagctcc ctctgagcca
     4861 ggagacattt tcaggcttat ggaaactact tcctccagaa gatatcctgc catcacctca
     4921 ctgcatggac gatctgttgc tgccccagga tgttgaggag ttttttgaag gcccaagtga
     4981 agccctccga gtgtcaggag ctcctgcagc acaggaccct gtcaccgaga cccctgggcc
     5041 agtggcccct gccccagcca ctccatggcc cctgtcatct tttgtccctt ctcaaaaaac
     5101 ttaccagggc aactatggct tccacctggg cttcctgcag tctgggacag ccaagtctgt
     5161 tatgtgcacg tactctcctc ccctcaataa gctattctgc cagctggcga agacgtgccc
     5221 tgtgcagttg tgggtcagcg ccacacctcc agctgggagc cgtgtccgcg ccatggccat
     5281 ctacaagaag tcacagcaca tgacggaggt cgtgagacgc tgcccccacc atgagcgctg
     5341 ctccgatggt gatggcctgg ctcctcccca gcatcttatc cgggtggaag gaaatttgta
     5401 tcccgagtat ctggaagaca ggcagacttt tcgccacagc gtggtggtac cttatgagcc
     5461 acccgaggcc ggctctgagt ataccaccat ccactacaag tacatgtgta atagctcctg
     5521 catggggggc atgaaccgcc gacctatcct taccatcatc acactggaag actccagtgg
     5581 gaaccttctg ggacgggaca gctttgaggt tcgtgtttgt gcctgccctg ggagagaccg
     5641 ccgtacagaa gaagaaaatt tccgcaaaaa ggaagtcctt tgccctgaac tgcccccagg
     5701 gagcgcaaag agagcgctgc ccacctgcac aagcgcctct cccccgcaaa agaaaaaacc
     5761 acttgatgga gagtatttca ccctcaagat ccgcgggcgt aaacgcttcg agatgttccg
     5821 ggagctaaat gaggccttag agttaaagga tgcccatgct acagaggagt ctggagacag
     5881 cagggctcac tccagctacc tgaagaccaa gaagggccag tctacttccc gccataaaaa
     5941 aacaatggtc aagaaagtgg ggcctgactc agactgatta attaagtcta gagtcgagtc
     6001 tagagggccc gcggttcgaa caaaaactca tctcagaaga ggatctgaat atgcataccg
     6061 gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta
     6121 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     6181 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     6241 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     6301 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     6361 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     6421 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     6481 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     6541 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     6601 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     6661 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     6721 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     6781 tgatttaaca aaaatttaac g
//
