LOCUS       sequence-167840-         832 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  hu6 driven sgRNA vector with G418 and GFP selectable markers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 832)
  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 832)
  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: pGH020_sgRNA_G418-GFP
FEATURES             Location/Qualifiers
     source          1..832
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     217..238
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      280..289
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(296..349)
                     /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"
     primer_bind     complement(467..486)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
       61 agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg
      121 gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct
      181 cgccctcgcc agacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
      241 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtgggc
      301 caggattctc ctcgacgtca ccgcatgtta gcagacttcc tctgccctct cccgatccga
      361 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
      421 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
      481 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
      541 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
      601 gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc
      661 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
      721 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
      781 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca ag
//
