LOCUS       sequence-167860-         788 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  hU6 driven sgRNA vector with 2xMS2 vectors with puro selectable 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 788)
  AUTHORS   Hess GT, Fresard L, Han K, Lee CH, Li A, Cimprich KA, Montgomery SB,
            Bassik MC
  TITLE     Directed evolution using dCas9-targeted somatic hypermutation in 
            mammalian cells.
  JOURNAL   Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4038.
  PUBMED    27798611
REFERENCE   2  (bases 1 to 788)
  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 
            Methods. 2016 Oct 31. doi: 10.1038/nmeth.4038."
COMMENT     Sequence Label: pGH224_sgRNA_2xMS2_Puro
FEATURES             Location/Qualifiers
     source          1..788
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(48..647)
                     /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(131..151)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     628..647
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(686..706)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 attttgtaat ccagaggttg attaccgata agcttgatat cgaattctca ggcaccgggc
       61 ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg
      121 aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg aggtctccag gaaggcgggc
      181 accccggcgc gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg
      241 ccctggtggt cgggcgatac gccgacggtg gccaggaacc acgcgggctc cttgggccgg
      301 tgcggcgcca ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg
      361 gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc
      421 cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg
      481 cgcgtgagga agagttcttg cagctcggtg acccgctcga tgtggcggtc cgggtcgacg
      541 gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggcccggggg
      601 acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact cggtcattgt acatcacgac
      661 acctgaaatg gaagaaaaaa actttgaacc actgtctgag gcttgagaat gaaccaagat
      721 ccaaactcaa aaagggcaaa ttccaaggag aattacatca agtgccaagc tggcctaact
      781 tcagtctc
//
