LOCUS       sequence-165593-         871 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Empty concatmer vector in which 4 gRNAs can be inserted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 871)
  AUTHORS   Andersson-Rolf A, Merenda A, Mustata RC, Li T, Dietmann S, Koo BK
  TITLE     Simultaneous paralogue knockout using a CRISPR-concatemer in mouse 
            small intestinal organoids.
  JOURNAL   Dev Biol. 2016 Oct 27. pii: S0012-1606(16)30479-1. doi: 
            10.1016/j.ydbio.2016.10.016.
  PUBMED    27983963
REFERENCE   2  (bases 1 to 871)
  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: "Dev Biol. 
            2016 Oct 27. pii: S0012-1606(16)30479-1. doi: 
            10.1016/j.ydbio.2016.10.016."
COMMENT     Sequence Label: 4 gRNA concatemer
FEATURES             Location/Qualifiers
     source          1..871
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        12..260
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        277..352
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             359..382
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        405..653
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        672..747
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             753..779
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             753..779
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
       61 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
      121 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      181 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      241 tgtggaaagg acgaaacacc gggtcttcga gaagacgttt tagagctaga aatagcaagt
      301 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttga
      361 ctacaaggac gacgatgaca aggatctcca aggtcgggca ggaagagggc ctatttccca
      421 tgattccttc atatttgcat atacgataca aggctgttag agagataatt agaattaatt
      481 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
      541 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
      601 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accggagtct
      661 tcgagaagac agttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
      721 cttgaaaaag tggcaccgag tcggtgcttt tttaccccta cgacgtgccc gactacgccg
      781 gatctccaag gtcgggcagg aagagggcct atttcccatg attccttcat atttgcatat
      841 acgatacaag gctgttagag agataattag a
//
