LOCUS       sequence-31780-e         851 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Entry vector for cloning miR30-based shRNA driven by TRE-tight 
            promoter with GFP coexpression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 851)
  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 851)
  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_TTGmiRc2
FEATURES             Location/Qualifiers
     source          1..851
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     ncRNA           49..170
                     /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             206..511
                     /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     390..409
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     ncRNA           532..658
                     /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(697..796)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 acgagctgta caagtaaact agtcgactag ggataacagg gtaattgttt gaatgaggct
       61 tcagtacttt acagaatcgt tgcctgcaca tcttggaaac acttgctggg attacttctt
      121 caggttaacc caacagaagg ctcgagaagg tatattgctg ttgacagtga gcgtgctgca
      181 ggtaagcttg aggtgtgctt ccagaatgca gtttaaggtt tacacctata aaagagagag
      241 ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg ggcgacggat
      301 agtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc
      361 ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc
      421 ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa
      481 cgccattaac ctgatgttct ggggaatata gaattccaac ctgctgcttg cctactgcct
      541 cggaattcaa ggggctactt taggagcaat tatcttgttt actaaaactg aataccttgc
      601 tatctctttg atacattttt acaaagctga attaaaatgg tataaattaa atcacttttt
      661 tcaattggaa gactaatgcg ttttcgagat atctagaccc agctttcttg tacaaagttg
      721 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
      781 ataaaatcat tatttgccat ccagctgcag ctctggcccg tgtctcaaaa tctctgatgt
      841 tacattgcac a
//
