LOCUS       sequence-31776-e         868 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Entry vector for cloning miR30-based shRNA driven by TRE promoter 
            with GFP coexpression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 868)
  AUTHORS   Shin KJ, Wall EA, Zavzavadjian JR, Santat LA, Liu J, Hwang JI, 
            Rebres R, Roach T, Seaman W, Simon MI, Fraser ID
  TITLE     A single lentiviral vector platform for microRNA-based conditional 
            RNA interference and coordinated transgene expression.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64.
  PUBMED    16945906
REFERENCE   2  (bases 1 to 868)
  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: "Proc Natl 
            Acad Sci U S A. 2006 Sep 12. 103(37):13759-64."
COMMENT     Sequence Label: pEN_TGmiRc3
FEATURES             Location/Qualifiers
     source          1..868
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     ncRNA           62..183
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     CDS             219..524
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRIVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     403..422
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     ncRNA           545..671
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     protein_bind    complement(710..809)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gagctgtaca agtaaactag tgactctagt cctagtcgac tagggataac agggtaattg
       61 tttgaatgag gcttcagtac tttacagaat cgttgcctgc acatcttgga aacacttgct
      121 gggattactt cttcaggtta acccaacaga aggctcgaga aggtatattg ctgttgacag
      181 tgagcgtgct gcaggtaagc ttgaggtgtg cttccagaat gcagtttaag gtttacacct
      241 ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc
      301 ccgggcgacg gatagtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc
      361 gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata
      421 tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa
      481 atgacatcaa aaacgccatt aacctgatgt tctggggaat atagaattcc aacctgctgc
      541 ttgcctactg cctcggaatt caaggggcta ctttaggagc aattatcttg tttactaaaa
      601 ctgaatacct tgctatctct ttgatacatt tttacaaagc tgaattaaaa tggtataaat
      661 taaatcactt ttttcaattg gaagactaat gcgttttcga gatatctaga cccagctttc
      721 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
      781 actatcagtc aaaataaaat cattatttgc catccagctg cagctctggc ccgtgtctca
      841 aaatctctga tgttacattg cacaagat
//
