LOCUS       sequence-166231-         865 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral expression of shRNAs, compatible with double-shRNA 
            expression, SFFV promoter, BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 865)
  AUTHORS   Kampmann M, Horlbeck MA, Chen Y, Tsai JC, Bassik MC, Gilbert LA, 
            Villalta JE, Kwon SC, Chang H, Kim VN, Weissman JS
  TITLE     Next-generation libraries for robust RNA interference-based 
            genome-wide screens.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Jun 30;112(26):E3384-91. doi: 
            10.1073/pnas.1508821112. Epub 2015 Jun 15.
  PUBMED    26080438
REFERENCE   2  (bases 1 to 865)
  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"; doi: 
            "10.1073/pnas.1508821112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-06-30- 30"; volume: "112"; issue: "26"; pages: 
            "E3384-91"
COMMENT     Sequence Label: pMK1224
FEATURES             Location/Qualifiers
     source          1..865
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             48..644
                     /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(48..67)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     544..564
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             654..707
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
ORIGIN
        1 ggcgcgccag tcctccgaca gactgagtcg cccgggtcta gcccaccatg accgagtaca
       61 agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg
      121 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc
      181 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
      241 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
      301 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
      361 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
      421 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
      481 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
      541 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
      601 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgccggatcg ggagagggca
      661 gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccagcg gccgccacca
      721 tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca
      781 accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
      841 tgagaatcaa ggtggtcgag ggcgg
//
