LOCUS       sequence-80468-e        1034 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 1034)
  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 1034)
  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..1034
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    78..102
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(114..189)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(210..458)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(268..287)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(438..458)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    576..693
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt gtcgacttaa cgcgccaccg
       61 gttagcgcta gctaatgcca actttgtaca agaaagctgg gtctagaaaa aaagcaccga
      121 ctcggtgcca ctttttcaag ttgataacgg actagcctta ttttaacttg ctatttctag
      181 ctctaaaaca tcctgtccct agtggccccg gtgtttcgtc ctttccacaa gatatataaa
      241 gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat
      301 aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact
      361 aatatctttg tgtttacagt caaattaatt ctaattatct ctctaacagc cttgtatcgt
      421 atatgcaaat atgaaggaat catgggaaat aggccctctt cctgcccgac cttggtaccg
      481 gatccagtcg actgaattgg ttcctttaaa gcctgctttt ttgtacactc gagccggcgc
      541 caaagtggat ctctgctgtc cctgtaataa acccgaaaat tttgaatttt tgtaatttgt
      601 ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca
      661 ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc catttgtctc
      721 aagatctaga attcttacta ctgcagctgc aggccctccc acacataacc agagggcagc
      781 aattcacgaa tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc
      841 ccaggatgca gatcgagaag cacctgtcgg caccgtccgc aggggctcaa gatgcccctg
      901 ttctcatttc cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg
      961 cccagcacca cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg
     1021 aagatgcggc cgtc
//
