LOCUS       sequence-473373-        4925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express p53 response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4925)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4925)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pNL [NlucP/p53-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        129..160
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      187..196
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             193..705
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             709..828
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    880..1001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(891..910)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     945..964
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1195..1552
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1195..1386
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1218..1238)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1403..1538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1465..1484
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1583..2620
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2599..2607
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    2644..2692
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2848..2865)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3019..3607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3099..3118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3807..4598,4599..4667))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4772..4820
                     /note="synthetic polyadenylation signal"
     misc_feature    4834..4925
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4874..4893
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctctaca gaacatgtct aagcatgctg tgccttgcct
       61 ggacttgcct ggccttgcct tgggctcgag gatatcaaga tctggcctcg gcggccaagc
      121 ttagacacta gagggtatat aatggaagct cgacttccag cttggcaatc cggtactgtt
      181 ggtaaagcca ccatggtctt cacactcgaa gatttcgttg gggactggcg acagacagcc
      241 ggctacaacc tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc
      301 ggggtgtccg taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc
      361 gacatccatg tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa
      421 aaaattttta aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat
      481 ggcacactgg taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat
      541 gaaggcatcg ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc
      601 aacaaaatta tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc
      661 atcaacggag tgaccggctg gcggctgtgc gaacgcattc tggcgaattc tcacggcttt
      721 ccgcctgagg ttgaagagca agccgccggt acattgccta tgtcctgcgc acaagaaagc
      781 ggtatggacc ggcacccagc cgcttgtgct tcagctcgca tcaacgtcta aggccgcgac
      841 tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
      901 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
      961 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1021 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     1081 acaaatgtgg taaaatcgat aaggatccgt ttgcgtattg ggcgctcttc cgctgatctg
     1141 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttagcta ccttctgagg
     1201 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     1261 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     1321 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     1381 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     1441 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     1501 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcga
     1561 ttcttctgac actagcgcca ccatgaagaa gcccgaactc accgctacca gcgttgaaaa
     1621 atttctcatc gagaagttcg acagtgtgag cgacctgatg cagttgtcgg agggcgaaga
     1681 gagccgagcc ttcagcttcg atgtcggcgg acgcggctat gtactgcggg tgaatagctg
     1741 cgctgatggc ttctacaaag accgctacgt gtaccgccac ttcgccagcg ctgcactacc
     1801 catccccgaa gtgttggaca tcggcgagtt cagcgagagc ctgacatact gcatcagtag
     1861 acgcgcccaa ggcgttactc tccaagacct ccccgaaaca gagctgcctg ctgtgttaca
     1921 gcctgtcgcc gaagctatgg atgctattgc cgccgccgac ctcagtcaaa ccagcggctt
     1981 cggcccattc gggccccaag gcatcggcca gtacacaacc tggcgggatt tcatttgcgc
     2041 cattgctgat ccccatgtct accactggca gaccgtgatg gacgacaccg tgtccgccag
     2101 cgtagctcaa gccctggacg aactgatgct gtgggccgaa gactgtcccg aggtgcgcca
     2161 cctcgtccat gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc gcatcaccgc
     2221 cgtaatcgac tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg ccaacatctt
     2281 cttctggcgg ccctggctgg cttgcatgga gcagcagact cgctacttcg agcgccggca
     2341 tcccgagctg gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg gcctggatca
     2401 gctctaccag agcctcgtgg acggcaactt cgacgatgct gcctgggctc aaggccgctg
     2461 cgatgccatc gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg ctcgccggag
     2521 cgcagccgta tggaccgacg gctgcgtcga ggtgctggcc gacagcggca accgccggcc
     2581 cagtacacga ccgcgcgcta aggaggtagg tcgagtttaa actctagaac cggtcatggc
     2641 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgttcgaact
     2701 agatgctgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
     2761 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga
     2821 caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2881 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2941 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3001 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3061 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3181 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3421 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3721 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagcgg
     3781 ccgcaaatgc taaaccactg cagtggttac cagtgcttga tcagtgaggc accgatctca
     3841 gcgatctgcc tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta gatcactacg
     3901 attcgtgagg gcttaccatc aggccccagc gcagcaatga tgccgcgaga gccgcgttca
     3961 ccggcccccg atttgtcagc aatgaaccag ccagcaggga gggccgagcg aagaagtggt
     4021 cctgctactt tgtccgcctc catccagtct atgagctgct gtcgtgatgc tagagtaaga
     4081 agttcgccag tgagtagttt ccgaagagtt gtggccattg ctactggcat cgtggtatca
     4141 cgctcgtcgt tcggtatggc ttcgttcaac tctggttccc agcggtcaag ccgggtcaca
     4201 tgatcaccca tattatgaag aaatgcagtc agctccttag ggcctccgat cgttgtcaga
     4261 agtaagttgg ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa ttctcttacc
     4321 gtcatgccat ccgtaagatg cttttccgtg accggcgagt actcaaccaa gtcgttttgt
     4381 gagtagtgta tacggcgacc aagctgctct tgcccggcgt ctatacggga caacaccgcg
     4441 ccacatagca gtactttgaa agtgctcatc atcgggaatc gttcttcggg gcggaaagac
     4501 tcaaggatct tgccgctatt gagatccagt tcgatatagc ccactcttgc acccagttga
     4561 tcttcagcat cttttacttt caccagcgtt tcggggtgtg caaaaacagg caagcaaaat
     4621 gccgcaaaga agggaatgag tgcgacacga aaatgttgga tgctcatact cgtccttttt
     4681 caatattatt gaagcattta tcagggttac tagtacgtct ctcaaggata agtaagtaat
     4741 attaaggtac gggaggtatt ggacaggccg caataaaata tctttatttt cattacatct
     4801 gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac
     4861 gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt
     4921 tctct
//
