LOCUS       sequence-146929-        5009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luciferase reporter for miRNA activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5009)
  AUTHORS   Eichelser C, Stuckrath I, Muller V, Milde-Langosch K, Wikman H, 
            Pantel K, Schwarzenbach H
  TITLE     Increased serum levels of circulating exosomal microRNA-373 in 
            receptor-negative breast cancer patients.
  JOURNAL   Oncotarget. 2014 Oct 30;5(20):9650-63.
  PUBMED    25333260
REFERENCE   2  (bases 1 to 5009)
  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: 
            "Oncotarget."; date: "2014-10-30"; volume: "5(20)"; pages: "9650-63"
COMMENT     Sequence Label: CD 44 pGL4basic
FEATURES             Location/Qualifiers
     source          1..5009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      861..870
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             867..2519
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2563..2684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2574..2593)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2628..2647
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2932..2949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3103..3691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3183..3202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3891..4682,4683..4751))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4856..4904
                     /note="synthetic polyadenylation signal"
     misc_feature    4918..5009
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4958..4977
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgaggg atgggcggat ggaaggatat
       61 ttaggaggat gaatgagcat gtgtgtggag agaggtgccc attcacactg gcttgaacac
      121 atgggttagc tgagccaaat gccagcccta tgacaggcca tcagtagctt tccctgagct
      181 gttctgccaa gaagctaaaa ttcattcaag ccatgtggac ttgttattga ggggaaaaag
      241 aatgagctct ccctctttcc acttggaaga ttcaccaact ccccacccct cactccccac
      301 tgtgggcacg gaggcactgc gccacccagg gcaagacctc gccctctctc cagctcctct
      361 cccaggatat ccaacatcct gtgaaaccca gagatcttgc tccagccgga ttcagagaaa
      421 tttagcggga aaggagaggc caaaggctga acccaatggt gcaaggtttt acggttcggt
      481 catcctctgt cctgacgccg cggggccagc gggagaagaa agccagtgca tctctgggcg
      541 caggggccag tggggctcgg aggcacaggc accccgcgac actccaggtt ccccgaccca
      601 cgtccctggc agccccgatt atttacagcc tcagcagagc acggggcggg ggcagagggg
      661 cccgcccggg agggctgcta cttcttaaaa cctctgcggg ctgcttagtc acagcccccc
      721 ttgcttgggt gtgtccttcg ctcgctccct ccctccgtct taggtcactg tttttcaacc
      781 tcgaatgaaa actgcagcca actttccgag gcagccttca ttgcccagcg gaagcttggc
      841 aatccggtac tgttggtaaa gccaccatgg aagatgccaa aaacattaag aagggcccag
      901 cgccattcta cccactcgaa gacgggaccg ccggcgagca gctgcacaaa gccatgaagc
      961 gctacgccct ggtgcccggc accatcgcct ttaccgacgc acatatcgag gtggacatta
     1021 cctacgccga gtacttcgag atgagcgttc ggctggcaga agctatgaag cgctatgggc
     1081 tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg
     1141 tgttgggtgc cctgttcatc ggtgtggctg tggccccagc taacgacatc tacaacgagc
     1201 gcgagctgct gaacagcatg ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag
     1261 ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat catacaaaag atcatcatca
     1321 tggatagcaa gaccgactac cagggcttcc aaagcatgta caccttcgtg acttcccatt
     1381 tgccacccgg cttcaacgag tacgacttcg tgcccgagag cttcgaccgg gacaaaacca
     1441 tcgccctgat catgaacagt agtggcagta ccggattgcc caagggcgta gccctaccgc
     1501 accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc catcttcggc aaccagatca
     1561 tccccgacac cgctatcctc agcgtggtgc catttcacca cggcttcggc atgttcacca
     1621 cgctgggcta cttgatctgc ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc
     1681 tattcttgcg cagcttgcaa gactataaga ttcaatctgc cctgctggtg cccacactat
     1741 ttagcttctt cgctaagagc actctcatcg acaagtacga cctaagcaac ttgcacgaga
     1801 tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc
     1861 acctaccagg catccgccag ggctacggcc tgacagaaac aaccagcgcc attctgatca
     1921 cccccgaagg ggacgacaag cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta
     1981 aggtggtgga cttggacacc ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg
     2041 tccgtggccc catgatcatg agcggctacg ttaacaaccc cgaggctaca aacgctctca
     2101 tcgacaagga cggctggctg cacagcggcg acatcgccta ctgggacgag gacgagcact
     2161 tcttcatcgt ggaccggctg aagagcctga tcaaatacaa gggctaccag gtagccccag
     2221 ccgaactgga gagcatcctg ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc
     2281 tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa
     2341 ccatgaccga gaaggagatc gtggactatg tggccagcca ggttacaacc gccaagaagc
     2401 tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg actgaccggc aagttggacg
     2461 cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg cggcaagatc gccgtgtaat
     2521 aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt
     2581 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2641 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2701 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     2761 tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag agccttcaac
     2821 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     2881 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac
     2941 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3001 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3061 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3121 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3181 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3241 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3301 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3361 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3421 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3481 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3541 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3601 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3661 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3721 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3781 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3841 tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc
     3901 ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat agtggcctga
     3961 ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc cagcgcagca
     4021 atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa ccagccagca
     4081 gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca gtctatgagc
     4141 tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag agttgtggcc
     4201 attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt caactctggt
     4261 tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc agtcagctcc
     4321 ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct catggtaatg
     4381 gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc
     4441 gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg
     4501 gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct catcatcggg
     4561 aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc cagttcgata
     4621 tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag cgtttcgggg
     4681 tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt
     4741 tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg ttactagtac
     4801 gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag gccgcaataa
     4861 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac
     4921 atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag
     4981 tgcaagtgca ggtgccagaa catttctct
//
