LOCUS       sequence-359624-        4556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P53 Firefly luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4556)
  AUTHORS   Sarrion-Perdigones A, Gonzalez Y, Venken KJT
  TITLE     Synthetic Assembly DNA Cloning of Multiplex Hextuple Luciferase 
            Reporter Plasmids.
  JOURNAL   Methods Mol Biol. 2022;2524:409-432. doi: 
            10.1007/978-1-0716-2453-1_32.
  PUBMED    35821490
REFERENCE   2  (bases 1 to 4556)
  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: "Methods Mol
            Biol."; date: "2022"; volume: "2524:409-432. doi"; pages: " 
            10.1007/978-1-0716-2453-1_32"
COMMENT     Sequence Label: 2xP53_RE::FLuc
FEATURES             Location/Qualifiers
     source          1..4556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    26..74
                     /note="synthetic polyadenylation signal"
     misc_feature    88..179
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     128..147
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        292..323
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             357..2009
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(423..441)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2017..2241
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2366..2954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2446..2465)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3069..3171
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3172..3987
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3172..3987
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3245..3264)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4101..4529
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4188..4207)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4398..4419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taacagatct cgtctcagtc aggagaataa aatatcttta ttttcattac atctgtgtgt
       61 tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca
      121 aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctctc
      181 gctgtcagga gtacagaaca tgtctaagca tgctgtgcct tgcctggact tgcctggcct
      241 tgccttccct cgaggatatc aaatctggcc tcggcggcca agcttagaca ctagagggta
      301 tataatggaa gctcgacttc cagcttggca atccggtact gttggtaaag ccaccaatgg
      361 aagacgccaa aaacataaag aaaggcccgg cgccattcta tccgctggaa gatggaaccg
      421 ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga acaattgctt
      481 ttacagatgc acatatcgag gtggacatca cttacgctga gtacttcgaa atgtccgttc
      541 ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca
      601 gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc ggagttgcag
      661 ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg ggcatttcgc
      721 agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa
      781 agctcccaat catccaaaaa attattatca tggattctaa aacggattac cagggatttc
      841 agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa tacgattttg
      901 tgccagagtc cttcgatagg gacaagacaa ttgcactgat catgaactcc tctggatcta
      961 ctggtctgcc taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg
     1021 ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc
     1081 cattccatca cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag
     1141 tcgtcttaat gtatagattt gaagaagagc tgtttctgag gagccttcag gattacaaga
     1201 ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc actctgattg
     1261 acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg
     1321 aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa ggatatgggc
     1381 tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg
     1441 tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc
     1501 tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg tccggttatg
     1561 taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag
     1621 acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg aagtctctga
     1681 ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg ctccaacacc
     1741 ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg
     1801 ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg
     1861 tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag
     1921 taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg
     1981 ccaagaaggg cggaaagatc gccgtgtaat gagcttctgt gccttctagt tgccagccat
     2041 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2101 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2161 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2221 gggatgcggt gggctctatg gcgctaatgt gagacgagat cttgacacgg ttatccacag
     2281 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2341 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2401 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2461 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2521 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2581 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2641 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2701 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2761 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2821 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2881 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2941 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3001 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3061 ttttgtgatg atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta
     3121 ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa aatgagccat
     3181 attcaacggg aaacgtcttg ctcaaggccg cgattaaatt ccaacatgga tgctgattta
     3241 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctaccgattg
     3301 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     3361 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     3421 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatcccaggg
     3481 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     3541 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     3601 gatcgcgtat ttcgactcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     3661 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     3721 aaacttttgc cattctcacc ggattcagta gtcactcatg gtgatttctc acttgataac
     3781 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     3841 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     3901 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     3961 catttgatgc tcgatgagtt tttctgattt tttttaaggc agttattggt gcccttaaac
     4021 gcctggttgc tacgcctgaa taagtgataa taagcggatg aatggcagaa attcgatgat
     4081 aagctgtcaa acatgatgtt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4141 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4201 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4261 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4321 tgatggttca cgtagtgggc cagcgccctg atagacggtt tttcgccctt tgacgttgga
     4381 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4441 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4501 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caattt
//
