LOCUS       sequence-109364-        6871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase expression driven by miR-34a promoter P3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6871)
  AUTHORS   Chang TC, Wentzel EA, Kent OA, Ramachandran K, Mullendore M, Lee KH,
            Feldmann G, Yamakuchi M, Ferlito M, Lowenstein CJ, Arking DE, Beer 
            MA, Maitra A, Mendell JT
  TITLE     Transactivation of miR-34a by p53 broadly influences gene expression
            and promotes apoptosis.
  JOURNAL   Mol Cell. 2007 Jun 8;26(5):745-52. Epub 2007 May 31.
  PUBMED    17540599
REFERENCE   2  (bases 1 to 6871)
  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: "Mol Cell.";
            date: "2007-06-8"; volume: "26(5)"; pages: "745-52. Epub 2007 May 
            31"
COMMENT     Sequence Label: pGL3-IRES-miR34a-P3
FEATURES             Location/Qualifiers
     source          1..6871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      419..428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1522..2089
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1683..1700)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1910..1929
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2135..2144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2141..3793
                     /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(2207..2225)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3834..3955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3845..3864)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3899..3918
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4203..4220)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4374..4962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4454..4473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5133..5924,5925..5993))
                     /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     5756..5775
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5994..6098)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6125..6580
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6212..6231)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6422..6443
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6711..6759
                     /note="synthetic polyadenylation signal"
     misc_feature    6773..6864
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6813..6832
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagctt ccctgtcctc ctcctaaaga
       61 atcgtcttct cccagtgcag gaggaatgag tcccccgttc tcaccccggg gagcggttgc
      121 ctcgtgagtc caaggagaat ccgccctttc gttttgcgca ggtggagatc tcgcgggccc
      181 agttcggagt cctgcaaact gtgagtgtgc gaggagggct tgtttctgat gaaaccattt
      241 ctgtcccgta acaaccctga gggcagaaac ccttacaaaa ggagccaccg agagatttgc
      301 tgtccgcttg atactggggc acggtggtcc tgggggcagc ttgtcccgga agggcgcggg
      361 gcgcactggc tgggatctat aatggttggg gcaggagggt gggtggggag ccccttctcc
      421 atggcggcgg gaggtcttcg gcgctgcccc tgcccgcgcc tccctgccca ccccaagggc
      481 gcccgaccgt gcgcgcaggg cagcgtggaa agggtgggaa atgcagggag cgcgccttgg
      541 gggctgcccc atgctccccc ctgtagctcc gcgctcagga cccgcgtccg actttgcgtc
      601 tctgggaacc tttaaggccg gtgagggggg ctgagccgcg gcccctcggc tgagccaagc
      661 cctgccccac gcaaggtccc cagcctggcg cacccggctc cccagcgccg cgcgccggcc
      721 atgtcccttc ccaccccagc cccccgggac ccgagagccg tgcggggccg agcctcgtgc
      781 cccgaggcac gccgagccaa gtcacccctg cagcccgcgg cgccggaact tgggggcgca
      841 gtgagtgggg gcagggcgcc gggccggcgg gcggtggccg tggctgggtg ccggttcctg
      901 gctttagaag tcctttcgca tcttgttgaa tccggggctg cattggccct ccgagctctc
      961 cggactggca acgtcgcaga gacaacaggt cctgacccgg gacagacccc acacccgcgt
     1021 ccaggacgtg ggtttcttcc ctcttccagg cgagaggccc tcggactggg cgtcgcggcg
     1081 agccccccgt ggtttctgtt tgcttttgta cggtccccct tccccccacc accctgtccc
     1141 ggccccaccc gggccttggt cccaccgcga aggggcgagg agtccccgga agggtcgcga
     1201 tggccgggga gtccgggacc tcggctctgg gtaggcgcgc cccgcgaccc agcggcggcg
     1261 tgggcgaggg gcgctgcgca aactacgcgc tctcgctggg gaaacgccag cccaggcggg
     1321 cggcggcggt gaaggggatg aggaccagga ccccgtgccc ggcgggactg gccccgcacc
     1381 ccacgccgcc cccctgcggt gcgcaccgca tctgcgccct aggccgaggc caggctccga
     1441 cccctccccc gcaccgacgt gattcggatc gcgcggtgct ggcgccgcct tcatctcgag
     1501 atctgacccc atgcatcgaa ttccgccccc cccccctaac gttactggcc gaagccgctt
     1561 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     1621 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     1681 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
     1741 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     1801 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     1861 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     1921 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     1981 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     2041 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataag atctgcgatc
     2101 taagtaagct tggcattccg gtactgttgg taaagccacc atggaagacg ccaaaaacat
     2161 aaagaaaggc ccggcgccat tctatccgct ggaagatgga accgctggag agcaactgca
     2221 taaggctatg aagagatacg ccctggttcc tggaacaatt gcttttacag atgcacatat
     2281 cgaggtggac atcacttacg ctgagtactt cgaaatgtcc gttcggttgg cagaagctat
     2341 gaaacgatat gggctgaata caaatcacag aatcgtcgta tgcagtgaaa actctcttca
     2401 attctttatg ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga
     2461 catttataat gaacgtgaat tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt
     2521 cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca
     2581 aaaaattatt atcatggatt ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt
     2641 cgtcacatct catctacctc ccggttttaa tgaatacgat tttgtgccag agtccttcga
     2701 tagggacaag acaattgcac tgatcatgaa ctcctctgga tctactggtc tgcctaaagg
     2761 tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg catgccagag atcctatttt
     2821 tggcaatcaa atcattccgg atactgcgat tttaagtgtt gttccattcc atcacggttt
     2881 tggaatgttt actacactcg gatatttgat atgtggattt cgagtcgtct taatgtatag
     2941 atttgaagaa gagctgtttc tgaggagcct tcaggattac aagattcaaa gtgcgctgct
     3001 ggtgccaacc ctattctcct tcttcgccaa aagcactctg attgacaaat acgatttatc
     3061 taatttacac gaaattgctt ctggtggcgc tcccctctct aaggaagtcg gggaagcggt
     3121 tgccaagagg ttccatctgc caggtatcag gcaaggatat gggctcactg agactacatc
     3181 agctattctg attacacccg agggggatga taaaccgggc gcggtcggta aagttgttcc
     3241 attttttgaa gcgaaggttg tggatctgga taccgggaaa acgctgggcg ttaatcaaag
     3301 aggcgaactg tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca atccggaagc
     3361 gaccaacgcc ttgattgaca aggatggatg gctacattct ggagacatag cttactggga
     3421 cgaagacgaa cacttcttca tcgttgaccg cctgaagtct ctgattaagt acaaaggcta
     3481 tcaggtggct cccgctgaat tggaatccat cttgctccaa caccccaaca tcttcgacgc
     3541 aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt cccgccgccg ttgttgtttt
     3601 ggagcacgga aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac
     3661 aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac
     3721 cggaaaactc gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa
     3781 gatcgccgtg taattctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata
     3841 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3901 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3961 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     4021 caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg atgcccttga
     4081 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     4141 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct
     4201 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4261 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4321 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4381 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4441 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4501 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4561 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4621 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4681 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4741 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4801 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4861 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4921 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4981 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5041 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5101 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5161 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5221 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5281 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5341 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5401 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5461 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5521 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5581 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5641 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5701 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5761 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5821 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5881 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5941 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6001 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6061 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6121 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     6181 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     6241 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     6301 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     6361 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     6421 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     6481 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     6541 tttaacgcga attttaacaa aatattaacg cttacaattt gccattcgcc attcaggctg
     6601 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gcccaagcta
     6661 ccatgataag taagtaatat taaggtacgg gaggtacttg gagcggccgc aataaaatat
     6721 ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta ctaacatacg
     6781 ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa
     6841 gtgcaggtgc cagaacattt ctctatcgat a
//
