LOCUS       sequence-31781-e         831 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Entry vector for cloning miR30-based shRNA driven by TRE-tight 
            promoter with dsRed2 coexpression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 831)
  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 831)
  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_TTRmiRc2
FEATURES             Location/Qualifiers
     source          1..831
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    34..90
                     /label=MCS
                     /note="multiple cloning site"
     ncRNA           137..258
                     /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             294..599
                     /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     478..497
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     ncRNA           620..746
                     /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(785..809)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gagcgcaccg agggccgcca ccacctgttc ctgagatctc gagctcaagc ttcgaattct
       61 gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataaactag tcgactaggg
      121 ataacagggt aattgtttga atgaggcttc agtactttac agaatcgttg cctgcacatc
      181 ttggaaacac ttgctgggat tacttcttca ggttaaccca acagaaggct cgagaaggta
      241 tattgctgtt gacagtgagc gtgctgcagg taagcttgag gtgtgcttcc agaatgcagt
      301 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
      361 atattattga cacgcccggg cgacggatag tgatccccct ggccagtgca cgtctgctgt
      421 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
      481 tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc
      541 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataga
      601 attccaacct gctgcttgcc tactgcctcg gaattcaagg ggctacttta ggagcaatta
      661 tcttgtttac taaaactgaa taccttgcta tctctttgat acatttttac aaagctgaat
      721 taaaatggta taaattaaat cacttttttc aattggaaga ctaatgcgtt ttcgagatat
      781 ctagacccag ctttcttgta caaagttggc attataagaa agcattgctt a
//
