LOCUS       sequence-116716-         555 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expresses a single gRNA and VP64-dCas9-VP64.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 555)
  AUTHORS   Gersbach C
  TITLE     Multiplex CRISPR/Cas9 Genome Engineering for Directing Myogenic 
            Cellular Differentiation in Primary Human Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 555)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLV hU6-gRNA(anti-sense)
            hUbC-VP64-dCas9-VP64-...
FEATURES             Location/Qualifiers
     source          1..555
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        104..352
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     104..124
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     275..294
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        369..444
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    455..488
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 cgtctgacgt ggcagcgctc gccgtgagga ggggggcgcc cgcgggaggc gccaaaaccc
       61 ggcgcggagg ccagatctgg agccgacacg ggttaattaa taagagggcc tatttcccat
      121 gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt
      181 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
      241 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
      301 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccggagacgg
      361 acgtctcagt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
      421 gaaaaagtgg caccgagtcg gtgctttttt tttaataact tcgtataatg tatgctatac
      481 gaagttatta attaaccaaa ctggatctct gctgtccctg taataaaccc gaaaattttg
      541 aatttttgta atttg
//
