LOCUS       sequence-31779-e         868 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Entry vector for cloning miR30-based shRNA driven by TRE-tight 
            promoter..
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_TTmiRc2
FEATURES             Location/Qualifiers
     source          1..868
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     ncRNA           1..122
                     /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             158..463
                     /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     342..361
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     ncRNA           484..610
                     /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(649..748)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ttgaatgagg cttcagtact ttacagaatc gttgcctgca catcttggaa acacttgctg
       61 ggattacttc ttcaggttaa cccaacagaa ggctcgagaa ggtatattgc tgttgacagt
      121 gagcgtgctg caggtaagct tgaggtgtgc ttccagaatg cagtttaagg tttacaccta
      181 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
      241 cgggcgacgg atagtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg
      301 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
      361 ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
      421 tgacatcaaa aacgccatta acctgatgtt ctggggaata tagaattcca acctgctgct
      481 tgcctactgc ctcggaattc aaggggctac tttaggagca attatcttgt ttactaaaac
      541 tgaatacctt gctatctctt tgatacattt ttacaaagct gaattaaaat ggtataaatt
      601 aaatcacttt tttcaattgg aagactaatg cgttttcgag atatctagac ccagctttct
      661 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
      721 ctatcagtca aaataaaatc attatttgcc atccagctgc agctctggcc cgtgtctcaa
      781 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
      841 gcttacataa acagtaatac aagggtgt
//
