LOCUS       sequence-109363-        7177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase expression driven by miR-34a promoter P2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7177)
  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 7177)
  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-P2
FEATURES             Location/Qualifiers
     source          1..7177
                     /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    1828..2395
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1989..2006)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2216..2235
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2441..2450
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2447..4099
                     /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(2513..2531)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4140..4261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4151..4170)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4205..4224
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4509..4526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4680..5268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4760..4779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5439..6230,6231..6299))
                     /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     6062..6081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6300..6404)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6431..6886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6518..6537)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6728..6749
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7017..7065
                     /note="synthetic polyadenylation signal"
     misc_feature    7079..7170
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7119..7138
                     /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 tcatgcgccc
     1501 tgcctggccc ccacctggtc ctctttcctt ttcaggtgga ggagatgccg ctgtcccgtc
     1561 ggtctgggga cagcccagct ccccggatcc cgggctggag agacgcgtcg cggccccggg
     1621 gcctggtggc acgagcagga aggaggaccc ggcggcgggc tctgcctggg cttgcctggg
     1681 cttgttccga gccgggctgc ttctcggtga ccacgcagat cgggggcatt tggagatttt
     1741 gcgggagtcc tgcagccaag ctccggggca ggagaggcct ggaagcctgc actacctgct
     1801 ctcgagatct gaccccatgc atcgaattcc gccccccccc cctaacgtta ctggccgaag
     1861 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     1921 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     1981 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     2041 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     2101 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     2161 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     2221 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     2281 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     2341 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagatct
     2401 gcgatctaag taagcttggc attccggtac tgttggtaaa gccaccatgg aagacgccaa
     2461 aaacataaag aaaggcccgg cgccattcta tccgctggaa gatggaaccg ctggagagca
     2521 actgcataag gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc
     2581 acatatcgag gtggacatca cttacgctga gtacttcgaa atgtccgttc ggttggcaga
     2641 agctatgaaa cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc
     2701 tcttcaattc tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc
     2761 gaacgacatt tataatgaac gtgaattgct caacagtatg ggcatttcgc agcctaccgt
     2821 ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat
     2881 catccaaaaa attattatca tggattctaa aacggattac cagggatttc agtcgatgta
     2941 cacgttcgtc acatctcatc tacctcccgg ttttaatgaa tacgattttg tgccagagtc
     3001 cttcgatagg gacaagacaa ttgcactgat catgaactcc tctggatcta ctggtctgcc
     3061 taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc
     3121 tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca
     3181 cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat
     3241 gtatagattt gaagaagagc tgtttctgag gagccttcag gattacaaga ttcaaagtgc
     3301 gctgctggtg ccaaccctat tctccttctt cgccaaaagc actctgattg acaaatacga
     3361 tttatctaat ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg aagtcgggga
     3421 agcggttgcc aagaggttcc atctgccagg tatcaggcaa ggatatgggc tcactgagac
     3481 tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt
     3541 tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa
     3601 tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg tccggttatg taaacaatcc
     3661 ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag acatagctta
     3721 ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa
     3781 aggctatcag gtggctcccg ctgaattgga atccatcttg ctccaacacc ccaacatctt
     3841 cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt
     3901 tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca
     3961 agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg
     4021 tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg
     4081 cggaaagatc gccgtgtaat tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     4141 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4201 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4261 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     4321 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc
     4381 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     4441 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     4501 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4561 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4621 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4681 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4741 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4801 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4861 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4921 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4981 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5041 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5101 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5161 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5221 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5281 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5341 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5401 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5461 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5521 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5581 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5641 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5701 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5761 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5821 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5881 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5941 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6001 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6061 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6121 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6181 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6241 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6301 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6361 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6421 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6481 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6541 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6601 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     6661 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     6721 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     6781 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     6841 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc
     6901 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc
     6961 aagctaccat gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata
     7021 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa
     7081 catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca
     7141 gtgcaagtgc aggtgccaga acatttctct atcgata
//
