LOCUS       sequence-32232-e         979 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 979)
  AUTHORS   Ebert MS, Neilson JR, Sharp PA
  TITLE     MicroRNA sponges: competitive inhibitors of small RNAs in mammalian 
            cells.
  JOURNAL   Nat Methods. 2007 Sep . 4(9):721-6.
  PUBMED    17694064
REFERENCE   2  (bases 1 to 979)
  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: "Nat 
            Methods. 2007 Sep . 4(9):721-6."
COMMENT     Sequence Label: CMV-d2eGFP-empty
FEATURES             Location/Qualifiers
     source          1..979
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     50..69
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        50..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      129..138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(135..137,138..140,141..851,852..857,858..977)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(180..201)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(441..460)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     788..809
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
       61 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      121 caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      181 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      241 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
      301 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
      361 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      421 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
      481 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
      541 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
      601 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
      661 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
      721 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
      781 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
      841 agctgtacaa gaagcttagc catggcttcc cgccggaggt ggaggagcag gatgatggca
      901 cgctgcccat gtcttgtgcc caggagagcg ggatggaccg tcaccctgca gcctgtgctt
      961 ctgctaggat caatgtgta
//
