LOCUS       sequence-125696-         799 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral expression of AAVS1-targeting sgRNA; insert can be 
            replaced with custom sgRNA (see protocol).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 799)
  AUTHORS   Wang T, Wei JJ, Sabatini DM, Lander ES
  TITLE     Genetic screens in human cells using the CRISPR-Cas9 system.
  JOURNAL   Science. 2014 Jan 3;343(6166):80-4. doi: 10.1126/science.1246981. 
            Epub 2013 Dec 12.
  PUBMED    24336569
REFERENCE   2  (bases 1 to 799)
  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: 
            "10.1126/science.1246981"; journalName: "Science"; date: "2014-01-3-
            3"; volume: "343"; issue: "6166"; pages: "80-4"
COMMENT     Sequence Label: pLX-sgRNA
FEATURES             Location/Qualifiers
     source          1..799
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    77..101
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(113..188)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(209..457)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(267..286)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(437..457)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    575..692
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 agtcaatctt tcacaaattt tgtaatccag aggttgattg tcgacttaac gcgccaccgg
       61 ttagcgctag ctaatgccaa ctttgtacaa gaaagctggg tctagaaaaa aagcaccgac
      121 tcggtgccac tttttcaagt tgataacgga ctagccttat tttaacttgc tatttctagc
      181 tctaaaacat cctgtcccta gtggccccgg tgtttcgtcc tttccacaag atatataaag
      241 ccaagaaatc gaaatacttt caagttacgg taagcatatg atagtccatt ttaaaacata
      301 attttaaaac tgcaaactac ccaagaaatt attactttct acgtcacgta ttttgtacta
      361 atatctttgt gtttacagtc aaattaattc taattatctc tctaacagcc ttgtatcgta
      421 tatgcaaata tgaaggaatc atgggaaata ggccctcttc ctgcccgacc ttggtaccgg
      481 atccagtcga ctgaattggt tcctttaaag cctgcttttt tgtacactcg agccggcgcc
      541 aaagtggatc tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt
      601 tttgtaattc tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac
      661 tgtacccccc aatcccccct tttcttttaa aattgtggat gaatactgcc atttgtctca
      721 agatctagaa ttcttactac tgcagctgca ggccctccca cacataacca gagggcagca
      781 attcacgaat cccaactgc
//
