LOCUS       sequence-166235-         818 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral expression of shRNAs, compatible with double-shRNA 
            expression, EF1alpha promoter, mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 818)
  AUTHORS   Kampmann M, Bassik MC, Weissman JS
  TITLE     Integrated platform for genome-wide screening and construction of 
            high-density genetic interaction maps in mammalian cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Jun 18;110(25):E2317-26. doi: 
            10.1073/pnas.1307002110. Epub 2013 Jun 5.
  PUBMED    23739767
REFERENCE   2  (bases 1 to 818)
  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.1307002110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-06-18- 18"; volume: "110"; issue: "25"; pages: 
            "E2317-26"
COMMENT     Sequence Label: pMK1200
FEATURES             Location/Qualifiers
     source          1..818
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             16..612
                     /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(16..35)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     512..532
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             622..675
                     /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"
     regulatory      682..691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gtgagctagc ccaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
       61 gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
      121 accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
      181 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
      241 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
      301 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
      361 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
      421 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
      481 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
      541 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
      601 aagcccggtg ccggatcggg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag
      661 gagaatcctg gcccaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc
      721 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
      781 ttcgagatcg agggcgaggg cgagggccgc ccctacga
//
