LOCUS       sequence-456475-        6066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luciferase reporter for human miR-34a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6066)
  AUTHORS   Li J, Wang K, Chen X, Meng H, Song M, Wang Y, Xu X, Bai Y
  TITLE     Transcriptional activation of microRNA-34a by NF-kappa B in human 
            esophageal cancer cells.
  JOURNAL   BMC Mol Biol. 2012 Jan 31;13:4.
  PUBMED    22292433
REFERENCE   2  (bases 1 to 6066)
  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: "BMC Mol 
            Biol."; date: "2012-01-31"; volume: "13"; pages: "4"
COMMENT     Sequence Label: miR-34aP1
FEATURES             Location/Qualifiers
     source          1..6066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1330..1339
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1336..2988
                     /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(1402..1420)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3029..3150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3040..3059)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3094..3113
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3398..3415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3569..4157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3649..3668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4328..5119,5120..5188))
                     /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     4951..4970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5189..5293)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5320..5775
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5407..5426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5617..5638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5906..5954
                     /note="synthetic polyadenylation signal"
     misc_feature    5968..6059
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6008..6027
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagtgc cggttcctgg ctttagaagt
       61 cctttcgcat cttgttgaat ccggggctgc attggccctc cgagctctcc ggactggcaa
      121 cgtcgcagag acaacaggtc ctgacccggg acagacccca cacccgcgtc caggacgtgg
      181 gtttcttccc tcttccaggc gagaggccct cggactgggc gtcgcggcga gccccccgtg
      241 gtttctgttt gcttttgtac ggtccccctt ccccccacca ccctgtcccg gccccacccg
      301 ggccttggtc ccaccgcgaa ggggcgagga gtccccggaa gggtcgcgat ggccggggag
      361 tccgggacct cggctctggg taggcgcgcc ccgcgaccca gcggcggcgt gggcgagggg
      421 cgctgcgcaa actacgcgct ctcgctgggg aaacgccagc ccaggcgggc ggcggcggtg
      481 aaggggatga ggaccaggac cccgtgcccg gcgggactgg ccccgcaccc cacgccgccc
      541 ccctgcggtg cgcaccgcat ctgcgcccta ggccgaggcc aggctccgac ccctcccccg
      601 caccgacgtg attcggatcg cgcggtgctg gcgccgcctt catgcgccct gcctggcccc
      661 cacctggtcc tctttccttt tcaggtggag gagatgccgc tgtcccgtcg gtctggggac
      721 agcccagctc cccggatccc gggctggaga gacgcgtcgc ggccccgggg cctggtggca
      781 cgagcaggaa ggaggacccg gcggcgggct ctgcctgggc ttgcctgggc ttgttccgag
      841 ccgggctgct tctcggtgac cacgcagatc gggggcattt ggagattttg cgggagtcct
      901 gcagccaagc tccggggcag gagaggcctg gaagcctgca ctacctgctc gccccgtccc
      961 agcatgcacc caggtaaacg cttgtgtttc tcagtccgtg cgaaagtttg caaagaagga
     1021 ggcgggaact agaccaacaa ctttaataat catagtcata ataatgaaaa ccctgttcaa
     1081 ttgtgtagcc tccgtaaggg gaagaatttc caagaagcaa caaaatgttc tttgatttca
     1141 taatctctgg ggagaaagga ttcctttttt ggggttttgc tcggctagtt ctttcttccc
     1201 tcctctatct ccagcgtggg gcgaggaagg ccgactggtt catgggtggc aaatggggac
     1261 cctggctctc ttgcgctgct ggccacggtg atattgcagc aagcttggca ttccggtact
     1321 gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc gccattctat
     1381 ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg
     1441 gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac ttacgctgag
     1501 tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat
     1561 cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg
     1621 ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc
     1681 aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa
     1741 attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat ggattctaaa
     1801 acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt
     1861 tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat tgcactgatc
     1921 atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc
     1981 tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact
     2041 gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat
     2101 ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttctgagg
     2161 agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc
     2221 gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggt
     2281 ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca tctgccaggt
     2341 atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac acccgagggg
     2401 gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat
     2461 ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt gagaggtcct
     2521 atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat
     2581 ggatggctac attctggaga catagcttac tgggacgaag acgaacactt cttcatcgtt
     2641 gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc tgaattggaa
     2701 tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct tcccgacgat
     2761 gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa
     2821 aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga
     2881 gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc
     2941 agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg
     3001 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     3061 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3121 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3181 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     3241 aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     3301 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     3361 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3421 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3481 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3541 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3601 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3661 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3721 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3781 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3841 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3901 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3961 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4021 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4081 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4141 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4201 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4261 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4321 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4381 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4441 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4501 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4561 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4621 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4681 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4741 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4801 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4861 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4921 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4981 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5041 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5101 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5161 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5221 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5281 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     5341 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5401 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5461 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5521 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5581 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5641 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5701 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5761 taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     5821 tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg
     5881 tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     5941 tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca
     6001 aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta
     6061 tcgata
//
