LOCUS       sequence-138995-        1058 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  optimized lenti sgRNA cloning backbone with MS2 loops at tetraloop 
            and stemloop 2 and EF1a-puro resistance marker. Contains BsmBI sites
            for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1058)
  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 1058)
  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: lenti sgRNA(MS2)_puro optimized backbone
FEATURES             Location/Qualifiers
     source          1..1058
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        6..254
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6..26
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     177..196
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        292..310
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        362..380
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    472..589
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        747..958
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 ccagagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
       61 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
      121 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
      181 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
      241 aaggacgaaa cacccgagac gggataccgt ctctgtttta gagctaggcc aacatgagga
      301 tcacccatgt ctgcagggcc tagcaagtta aaataaggct agtccgttat caacttggcc
      361 aacatgagga tcacccatgt ctgcagggcc aagtggcacc gagtcggtgc ttttttggat
      421 ccaagcttgg cgtaactaga tcttgagaca aatggcagta ttcatccaca attttaaaag
      481 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
      541 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
      601 cagggacagc agagatccac tttggcgccg gctcgagggg gcccgggtgc aaagatggat
      661 aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt
      721 gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
      781 gttgggggga ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg
      841 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
      901 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt
      961 aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc
     1021 ttgaattact tccacctggc tgcagtacgt gattcttg
//
