LOCUS       sequence-109365-        5933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase expression driven by miR-34a promoter P4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5933)
  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 5933)
  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-P4
FEATURES             Location/Qualifiers
     source          1..5933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    584..1151
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(745..762)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     972..991
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      1197..1206
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1203..2855
                     /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(1269..1287)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2896..3017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2907..2926)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2961..2980
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3265..3282)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3436..4024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3516..3535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4195..4986,4987..5055))
                     /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     4818..4837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5056..5160)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5187..5642
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5274..5293)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5484..5505
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5773..5821
                     /note="synthetic polyadenylation signal"
     misc_feature    5835..5926
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5875..5894
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagcgc cctgcctggc ccccacctgg
       61 tcctctttcc ttttcaggtg gaggagatgc cgctgtcccg tcggtctggg gacagcccag
      121 ctccccggat cccgggctgg agagacgcgt cgcggccccg gggcctggtg gcacgagcag
      181 gaaggaggac ccggcggcgg gctctgcctg ggcttgcctg ggcttgttcc gagccgggct
      241 gcttctcggt gaccacgcag atcgggggca tttggagatt ttgcgggagt cctgcagcca
      301 agctccgggg caggagaggc ctggaagcct gcactacctg ctcgccccgt cccagcatgc
      361 acccaggtaa acgcttgtgt ttctcagtcc gtgcgaaagt ttgcaaagaa ggaggcggga
      421 actagaccaa caactttaat aatcatagtc ataataatga aaaccctgtt caattgtgta
      481 gcctccgtaa ggggaagaat ttccaagaag caacaaaatg ttctttgatt tcataatctc
      541 tggggagaaa ggattcctcg agatctgacc ccatgcatcg aattccgccc ccccccccta
      601 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
      661 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
      721 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
      781 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
      841 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
      901 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
      961 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     1021 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     1081 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     1141 cacgatgata agatctgcga tctaagtaag cttggcattc cggtactgtt ggtaaagcca
     1201 ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg
     1261 gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa
     1321 ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt
     1381 ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg
     1441 tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag
     1501 ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca
     1561 tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc
     1621 aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg
     1681 gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg
     1741 attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg
     1801 gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct
     1861 cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg
     1921 ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat
     1981 ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt
     2041 acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc
     2101 tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct
     2161 ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat
     2221 atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg
     2281 gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga
     2341 aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg
     2401 gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt
     2461 ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt
     2521 ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc
     2581 aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac
     2641 ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg
     2701 attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg
     2761 acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca
     2821 taaaggccaa gaagggcgga aagatcgccg tgtaattcta gagtcggggc ggccggccgc
     2881 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2941 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3001 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3061 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     3121 atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     3181 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     3241 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3301 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3361 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3421 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3481 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3541 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3601 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3661 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3721 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3781 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3841 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3901 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3961 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4021 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4081 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4141 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4801 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5161 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5221 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5281 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5341 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5401 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5461 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5521 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5581 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5641 ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5701 gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact
     5761 tggagcggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     5821 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     5881 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctatcg ata
//
