LOCUS       sequence-95098-e         986 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd gen lentiviral backbone for cloning and expression of new shRNA 
            sequences. Uses puromycin for selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 986)
  AUTHORS   Stewart SA, Dykxhoorn DM, Palliser D, Mizuno H, Yu EY, An DS, 
            Sabatini DM, Chen IS, Hahn WC, Sharp PA, Weinberg RA, Novina CD
  TITLE     Lentivirus-delivered stable gene silencing by RNAi in primary cells.
  JOURNAL   RNA 2003 Apr;9(4):493-501.
  PUBMED    12649500
REFERENCE   2  (bases 1 to 986)
  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: "RNA"; date:
            "2003-04"; volume: "9(4)"; pages: "493-501"
COMMENT     Sequence Label: pLKO.1 puro
FEATURES             Location/Qualifiers
     source          1..986
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             70..669
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(70..89)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     566..586
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 tcggcagtcg gctccctcgt tgaccgaatc accgacctct ctccccaggg ggatccaccg
       61 gagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
      121 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
      181 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
      241 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
      301 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
      361 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
      421 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
      481 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
      541 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
      601 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
      661 ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat
      721 gcatcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
      781 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
      841 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
      901 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
      961 cgtctgttgt gtgactctgg taacta
//
