LOCUS       sequence-429525-        4676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human P53DD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4676)
  AUTHORS   Wu Y, Zhong A, Sidharta M, Kim TW, Ramirez B, Persily B, Studer L, 
            Zhou T
  TITLE     Robust and inducible genome editing via an all-in-one prime editor 
            in human pluripotent stem cells.
  JOURNAL   Nat Commun. 2024 Dec 30;15(1):10824. doi: 
            10.1038/s41467-024-55104-1.
  PUBMED    39737975
REFERENCE   2  (bases 1 to 4676)
  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"; journalName: "Nat 
            Commun."; date: "2024-12-30"; volume: "15(1):10824. doi"; pages: " 
            10.1038/s41467-024-55104-1"
COMMENT     Sequence Label: EF1a-hP53DD
FEATURES             Location/Qualifiers
     source          1..4676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        377..481
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(482..550,551..1342)
                     /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(700..719)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1513..2101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2002..2021
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2255..2272
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2340..2474
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2364..2383)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2418..2437
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(2480..2675)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2494..2629
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    2734..2755
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2770..2775,2776..2793,2794..2800)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2808..2824
                     /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     2813..2835
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2832..2848
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2832..2848
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3191..3210
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    3209..3264
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3310..3329
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     regulatory      3754..3763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(join(3809..4676,1..315))
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(3818..4676,1..84)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(3841..3861)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 caggggcgtg gaagtaattc aaggcacgca agggccataa cccgtaaaga ggccaggccc
       61 gcgggaacca cacacggcac ttacctgtgt tctggcggca aacccgttgc gaaaaagaac
      121 gttcacggcg actactgcac ttatatacgg ttctccccca ccctcgggaa aaaggcggag
      181 ccagtacacg acatcacttt cccagtttac cccgcgccac cttctctagg caccggttca
      241 attgccgacc cctcccccca acttctcggg gactgtgggc gatgtgcgct ctgcccactg
      301 acgggcaccg gagcctcacg cgatgatcta gtcaataatc aatgtcgacc caggtggcac
      361 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      421 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      481 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      541 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
      601 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
      661 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
      721 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
      781 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
      841 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
      901 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
      961 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1021 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1081 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     1141 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     1201 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     1261 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1321 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1381 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1441 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1501 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1561 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1621 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1681 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1741 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1801 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     1861 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     1921 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     1981 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2041 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2101 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2161 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2221 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     2281 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcgg
     2341 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2401 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2461 gcaataaaca agttagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     2521 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     2581 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     2641 ggggcgggac tatggttgct gactaattga gatgcggatc cgctggcacg acaggtttcc
     2701 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     2761 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     2821 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgggc tgcaggtcga
     2881 gggatctcca taagagaaga gggacagcta tgactgggag tagtcaggag aggaggaaaa
     2941 atctggctag taaaacatgt aaggaaaatt ttagggatgt taaagaaaaa aataacacaa
     3001 aacaaaatat aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga
     3061 cagcaggggg ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag
     3121 ccagcatatg ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat
     3181 gatttgccct cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg
     3241 caatgaaaat aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt
     3301 ccccataatt tttggcagag ggaaaaagat ctcagtggta tttgtgagcc agggcattgg
     3361 ccacaccagc caccaccttc tgataggcag cctgcacctg aggagtgcgg ccgctagcgg
     3421 gtttaaacaa cttggcgcgc ctaaacaccg gtcagtctga gtcaggccct tctgtcttga
     3481 acatgagttt tttatggcgg gaggtagact gacccttttt ggacttcagg tggctggagt
     3541 gagccctgct cccccctggc tccttcccag cctgggcatc cttgagttcc aaggcctcat
     3601 tcagctctcg gaacatctcg aagcgctcac gcccacggat ctgaagggtg aaatattctc
     3661 catccagtgg tttcttcttt ggctggggag aggagctggt gttgttgggc agtgctcgct
     3721 tagtgctccc tgggggcagc tcgtggtgag gctccatggt ggcgaccggt ggatcccggg
     3781 cccgcggtac cgtcgactgc agaattcctc acgacacctg aaatggaaga aaaaaacttt
     3841 gaaccactgt ctgaggcttg agaatgaacc aagatccaaa ctcaaaaagg gcaaattcca
     3901 aggagaatta catcaagtgc caagctggcc taacttcagt ctccacccac tcagtgtggg
     3961 gaaactccat cgcataaaac ccctcccccc aacctaaaga cgacgtactc caaaagctcg
     4021 agaactaatc gaggtgcctg gacggcgccc ggtactccgt ggagtcacat gaagcgacgg
     4081 ctgaggacgg aaaggccctt ttcctttgtg tgggtgactc acccgcccgc tctcccgagc
     4141 gccgcgtcct ccattttgag ctccctgcag cagggccggg aagcggccat ctttccgctc
     4201 acgcaactgg tgccgaccgg gccagccttg ccgcccaggg cggggcgata cacggcggcg
     4261 cgaggccagg caccagagca ggccggccag cttgagacta cccccgtccg attctcggtg
     4321 gccgcgctcg caggccccgc ctcgccgaac atgtgcgctg ggacgcacgg gccccgtcgc
     4381 cgcccgcggc cccaaaaacc gaaataccag tgtgcagatc ttggcccgca tttacaagac
     4441 tatcttgcca gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg gctagagact
     4501 tatcgaaagc agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg cggccccagc
     4561 gcccaggcca ggcctcaact caagcacgag gcgaaggggc tccttaagcg caaggcctcg
     4621 aactctccca cccacttcca acccgaagct cgggatcaag aatcacgtac tgcagc
//
