LOCUS       sequence-109367-        5626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase expression driven by miR-34a promoter P6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5626)
  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 5626)
  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-P6
FEATURES             Location/Qualifiers
     source          1..5626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    277..844
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(438..455)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     665..684
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      890..899
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             896..2548
                     /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(962..980)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2589..2710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2600..2619)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2654..2673
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2958..2975)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3129..3717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3209..3228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3888..4679,4680..4748))
                     /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     4511..4530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4749..4853)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4880..5335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4967..4986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5177..5198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5466..5514
                     /note="synthetic polyadenylation signal"
     misc_feature    5528..5619
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5568..5587
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagccc cgtcccagca tgcacccagg
       61 taaacgcttg tgtttctcag tccgtgcgaa agtttgcaaa gaaggaggcg ggaactagac
      121 caacaacttt aataatcata gtcataataa tgaaaaccct gttcaattgt gtagcctccg
      181 taaggggaag aatttccaag aagcaacaaa atgttctttg atttcataat ctctggggag
      241 aaaggattcc tcgagatctg accccatgca tcgaattccg cccccccccc ctaacgttac
      301 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
      361 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
      421 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
      481 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
      541 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
      601 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
      661 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
      721 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
      781 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
      841 ataagatctg cgatctaagt aagcttggca ttccggtact gttggtaaag ccaccatgga
      901 agacgccaaa aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc
      961 tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt
     1021 tacagatgca catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg
     1081 gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag
     1141 tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt
     1201 tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca
     1261 gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa
     1321 gctcccaatc atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca
     1381 gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt
     1441 gccagagtcc ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac
     1501 tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc
     1561 cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc
     1621 attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt
     1681 cgtcttaatg tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat
     1741 tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga
     1801 caaatacgat ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga
     1861 agtcggggaa gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct
     1921 cactgagact acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt
     1981 cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct
     2041 gggcgttaat caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt
     2101 aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga
     2161 catagcttac tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat
     2221 taagtacaaa ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc
     2281 caacatcttc gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc
     2341 cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt
     2401 cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt
     2461 accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc
     2521 caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg ggcggccggc cgcttcgagc
     2581 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2641 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2701 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2761 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc
     2821 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     2881 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     2941 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3001 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3061 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3121 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3181 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3241 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3301 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3361 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3421 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3481 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3541 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3601 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3661 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3721 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3781 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3841 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3901 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3961 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4021 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4081 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4141 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4201 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4261 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4321 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4381 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4441 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4501 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4561 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4621 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4681 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4741 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4801 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4861 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4921 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4981 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5041 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5101 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5161 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5221 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5281 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat
     5341 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     5401 cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg
     5461 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     5521 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
     5581 ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgata
//
