LOCUS       sequence-387844-        6801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of 3xHA tagged p53 L25Q,Q26S,F53Q,F54S.
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 QM
FEATURES             Location/Qualifiers
     source          1..6801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..25)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     252..272
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(281..709)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(391..412)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     603..622
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    755..979
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     968..985
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1008..1025)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1041..1070)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1900..1911
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(2308..2334)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(2338..2364)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(2368..2394)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2406..2415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2493..2512)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2494..2512)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2557..2760)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2586..2606)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2761..3140
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3312..3331
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3406..3510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3511..3579,3580..4371)
                     /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     complement(3729..3748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4542..5130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5031..5050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5284..5301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5418..5439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5454..5459,5460..5477,5478..5484)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5492..5508
                     /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)."
     primer_bind     5497..5519
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5516..5532
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5516..5532
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5581..5702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5592..5611)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5646..5665
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5878..6672)
                     /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(5989..6008)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6599..6618
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6739..6801,1..267))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6753..6801,1..87)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
       61 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
      121 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
      181 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
      241 ggactttcca caccctaact gacacacatt ccacagaatt aattcgcgtt aaatttttgt
      301 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
      361 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
      421 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
      481 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
      541 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
      601 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
      661 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg gggatacccc
      721 ctagagcccc agctggttct ttccgcctca gaagccatag agcccaccgc atccccagca
      781 tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
      841 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
      901 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
      961 tggcaactag aaggcacagt cgaggctgat cagcgggttt aaactcaatg gtgatggtga
     1021 tgatgaccgg tatgcatatt cagatcctct tctgagatga gtttttgttc gaaccgcggg
     1081 ccctctagac tcgactctag acttaattaa tcagtctgag tcaggcccca ctttcttgac
     1141 cattgttttt ttatggcggg aagtagactg gcccttcttg gtcttcaggt agctggagtg
     1201 agccctgctg tctccagact cctctgtagc atgggcatcc tttaactcta aggcctcatt
     1261 cagctcccgg aacatctcga agcgtttacg cccgcggatc ttgagggtga aatactctcc
     1321 atcaagtggt tttttctttt gcgggggaga ggcgcttgtg caggtgggca gcgctctctt
     1381 tgcgctccct gggggcagtt cagggcaaag gacttccttt ttgcggaaat tttcttcttc
     1441 tgtacggcgg tctctcccag ggcaggcaca aacacgaacc tcaaagctgt cccgtcccag
     1501 aaggttccca ctggagtctt ccagtgtgat gatggtaagg ataggtcggc ggttcatgcc
     1561 ccccatgcag gagctattac acatgtactt gtagtggatg gtggtatact cagagccggc
     1621 ctcgggtggc tcataaggta ccaccacgct gtggcgaaaa gtctgcctgt cttccagata
     1681 ctcgggatac aaatttcctt ccacccggat aagatgctgg ggaggagcca ggccatcacc
     1741 atcggagcag cgctcatggt gggggcagcg tctcacgacc tccgtcatgt gctgtgactt
     1801 cttgtagatg gccatggcgc ggacacggct cccagctgga ggtgtggcgc tgacccacaa
     1861 ctgcacaggg cacgtcttcg ccagctggca gaatagctta ttgaggggag gagagtacgt
     1921 gcacataaca gacttggctg tcccagactg caggaagccc aggtggaagc catagttgcc
     1981 ctggtaagtt ttttgagaag ggacaaaaga tgacaggggc catggagtgg ctggggcagg
     2041 ggccactggc ccaggggtct cggtgacagg gtcctgtgct gcaggagctc ctgacactcg
     2101 gagggcttca cttgggcctt cagactgctc ctcaacatcc tggggcagca acagatcgtc
     2161 catgcagtga ggtgatggca ggatatcttc tggaggaagt agtttcgatt ggcctgaaaa
     2221 tgtctcctgg ctcagaggga gctcgaggct gatatccgac tgtgactcct ccatggcagt
     2281 ggcgcgcccc cgcggccgca ctccttgagc gtagtctggg acgtcgtatg ggtagccagc
     2341 gtagtctggg acgtcgtatg ggtagccagc gtaatccgga acatcatacg ggtatccttg
     2401 ttgccccatg gtggcgttta tctgcagaat tccaccacac tggactagtg gatccgagct
     2461 cggtaccaag cttaactagc cagcttgggt ctccctatag tgagtcgtat taatttcgat
     2521 aagccagtaa gcagtgggtt ctctagttag ccagagagct ctgcttatat agacctccca
     2581 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     2641 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     2701 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     2761 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     2821 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     2881 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     2941 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     3001 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     3061 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     3121 actagtcaat aatcaatgtc aacgcgtata tctggcccgt acatcgcgaa gcagcgcaaa
     3181 acgcctaacc ctaagcagat tcttcatgca attgtcggtc aagccttgcc ttgttgtagc
     3241 ttaaattttg ctcgcgcact actcagcgac ctccaacaca caagcaggga gcagatactg
     3301 gcttaactat gcggcatcag agcagattgt actgagagtg caccataggg gatcgggaga
     3361 tctcccgatc cgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3421 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3481 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3541 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3601 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3661 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3721 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3781 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3841 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3901 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3961 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4021 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4081 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4141 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4201 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4261 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4321 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4381 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4441 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4681 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5161 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5221 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5281 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5341 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5401 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5461 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     5521 aacagctatg accatgatta cgccaagctc tagctagagg tcgacggtat acagacatga
     5581 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     5641 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     5701 ttggggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     5761 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     5821 cgtgatggca ggttgggcgt cgcttggtcg gtcatttcgc gaaccccaga gtcccgctca
     5881 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     5941 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     6001 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     6061 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     6121 gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag
     6181 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     6241 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     6301 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     6361 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     6421 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     6481 tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     6541 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     6601 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     6661 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatccga aaatggatat
     6721 acaagctccc gggagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     6781 ctggaatagc tcagaggcag a
//
