LOCUS       sequence-138990-         951 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation 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 951)
  AUTHORS   Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena 
            C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F
  TITLE     Genome-scale transcriptional activation by an engineered CRISPR-Cas9
            complex.
  JOURNAL   Nature. 2014 Dec 10. doi: 10.1038/nature14136.
  PUBMED    25494202
REFERENCE   2  (bases 1 to 951)
  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: "Nature. 
            2014 Dec 10. doi: 10.1038/nature14136."
COMMENT     Sequence Label: lenti sgRNA(MS2)_puro backbone
FEATURES             Location/Qualifiers
     source          1..951
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    126..243
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        293..541
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     293..313
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     464..483
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        579..597
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        649..667
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
ORIGIN
        1 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
       61 gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc
      121 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
      181 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
      241 tttcgggttt attacaggga cagcagagat ccagtttggt taattagcta gcgagggcct
      301 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattgg
      361 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
      421 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
      481 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
      541 cggagacggg ataccgtctc tgttttagag ctaggccaac atgaggatca cccatgtctg
      601 cagggcctag caagttaaaa taaggctagt ccgttatcaa cttggccaac atgaggatca
      661 cccatgtctg cagggccaag tggcaccgag tcggtgcttt ttttggatcc tgcaaagatg
      721 gataaagttt taaacagaga ggaatctttg cagctaatgg accttctagg tcttgaaagg
      781 agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
      841 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa
      901 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg g
//
