LOCUS       sequence-31777-e         920 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Entry vector for cloning miR30-based shRNA driven by TRE promoter 
            with dsRed2 coexpression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 920)
  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 920)
  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_TRmiRc2
FEATURES             Location/Qualifiers
     source          1..920
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    40..96
                     /label=MCS
                     /note="multiple cloning site"
     ncRNA           143..264
                     /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             300..605
                     /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     484..503
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     ncRNA           626..752
                     /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(791..890)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 cagtacgagc gcaccgaggg ccgccaccac ctgttcctga gatctcgagc tcaagcttcg
       61 aattctgcag tcgacggtac cgcgggcccg ggatccaccg gatctagata aactagtcga
      121 ctagggataa cagggtaatt gtttgaatga ggcttcagta ctttacagaa tcgttgcctg
      181 cacatcttgg aaacacttgc tgggattact tcttcaggtt aacccaacag aaggctcgag
      241 aaggtatatt gctgttgaca gtgagcgtgc tgcaggtaag cttgaggtgt gcttccagaa
      301 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
      361 agagtgatat tattgacacg cccgggcgac ggatagtgat ccccctggcc agtgcacgtc
      421 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
      481 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
      541 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
      601 tatagaattc caacctgctg cttgcctact gcctcggaat tcaaggggct actttaggag
      661 caattatctt gtttactaaa actgaatacc ttgctatctc tttgatacat ttttacaaag
      721 ctgaattaaa atggtataaa ttaaatcact tttttcaatt ggaagactaa tgcgttttcg
      781 agatatctag acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
      841 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
      901 gcagctctgg cccgtgtctc
//
