LOCUS       sequence-125401-        1051 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Retroviral transfer plasmid for Tet-regulated shRNA expression with 
            a TREtight promoter. Contains DsRed2 and Venus fluorescent 
            reporters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1051)
  AUTHORS   Zuber J, McJunkin K, Fellmann C, Dow LE, Taylor MJ, Hannon GJ, Lowe 
            SW
  TITLE     Toolkit for evaluating genes required for proliferation and survival
            using tetracycline-regulated RNAi.
  JOURNAL   Nat Biotechnol. 2011 Jan . 29(1):79-83.
  PUBMED    21131983
REFERENCE   2  (bases 1 to 1051)
  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: "Nat 
            Biotechnol. 2011 Jan . 29(1):79-83."
COMMENT     Sequence Label: TtRMPVIR
FEATURES             Location/Qualifiers
     source          1..1051
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        18..332
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    26..44
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    62..80
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    97..115
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    133..151
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    169..187
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    204..222
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    240..258
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     305..329
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             371..1048
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(551..571)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     963..986
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
ORIGIN
        1 gggggatcga taccgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta
       61 ctccctatca gtgatagaga acgatgtcga gtttactccc tatcagtgat agagaacgta
      121 tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
      181 atagagaacg tatgtcgagt ttatccctat cagtgataga gaacgtatgt cgagtttact
      241 ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt acggtgggag gcctatataa
      301 gcagagctcg tttagtgaac cgtcagatcg cctggagaat taattcgagc tcggtacccg
      361 gggatccacc atggcctcct ccgagaacgt catcaccgag ttcatgcgct tcaaggtgcg
      421 catggagggc accgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
      481 ctacgagggc cacaacaccg tgaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
      541 ctgggacatc ctgtcccccc agttccagta cggctccaag gtgtacgtga agcaccccgc
      601 cgacatcccc gactacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
      661 gaacttcgag gacggcggcg tggcgaccgt gacccaggac tcctccctgc aggacggctg
      721 cttcatctac aaggtgaagt tcatcggcgt gaacttcccc tccgacggcc ccgtgatgca
      781 gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct
      841 gaagggcgag acccacaagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
      901 caagtccatc tacatggcca agaagcccgt gcagctgccc ggctactact acgtggacgc
      961 caagctggac atcacctccc acaacgagga ctacaccatc gtggagcagt acgagcgcac
     1021 cgagggccgc caccacctgt tcctgtagcg g
//
