LOCUS       Exported                 788 bp ds-DNA     linear   SYN 17-MAY-2021
DEFINITION  Expression of a catalytically inactive, human codon-optimized Cas9 
            under the control of Murine Stem Cell retroVirus LTR promoter for 
            mammalian gene knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    pdCas9-humanized
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 788)
  AUTHORS   Qi LS, Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim 
            WA
  TITLE     Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific 
            Control of Gene Expression.
  JOURNAL   Cell. 2013 Feb 28;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022.
  PUBMED    23452860
REFERENCE   2  (bases 1 to 788)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 17, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..788
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             36..635
                     /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(36..55)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     532..552
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc
       61 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
      121 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
      181 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
      241 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
      301 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
      361 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
      421 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
      481 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
      541 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
      601 gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc
      661 cgaccgaaag gagcgcacga ccccatgcat cgataaaata aaagatttta tttagtctcc
      721 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
      781 cattttgc
//
