LOCUS       sequence-145679-         851 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  lenti sgRNA cloning backbone with MS2 loops at tetraloop and 
            stemloop 2, dCas9-VP64, and blast resistance marker. Contains BsmBI 
            sites for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 851)
  AUTHORS   Joung J, Konermann S, Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham
            MD, Sanjana NE, Zhang F
  TITLE     Genome-scale CRISPR-Cas9 knockout and transcriptional activation 
            screening.
  JOURNAL   Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016. 
            Epub 2017 Mar 23.
  PUBMED    28333914
REFERENCE   2  (bases 1 to 851)
  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: 
            "10.1038/nprot.2017.016"; journalName: "Nat Protoc"; date: 
            "2017-04"; volume: "12"; issue: "4"; pages: "828-863"
COMMENT     Sequence Label: lentiSAMv2
FEATURES             Location/Qualifiers
     source          1..851
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    113..230
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        280..528
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     280..300
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     451..470
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        566..584
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        636..654
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
ORIGIN
        1 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
       61 atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa
      121 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
      181 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
      241 acagggacag cagagatcca gtttggttaa ttagctagcg agggcctatt tcccatgatt
      301 ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat taatttgact
      361 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
      421 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
      481 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg agacgggata
      541 ccgtctctgt tttagagcta ggccaacatg aggatcaccc atgtctgcag ggcctagcaa
      601 gttaaaataa ggctagtccg ttatcaactt ggccaacatg aggatcaccc atgtctgcag
      661 ggccaagtgg caccgagtcg gtgctttttt tggatcctag gtcttgaaag gagtgggaat
      721 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      781 gggaggggtc ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      841 tgatgtcgtg t
//
