LOCUS       sequence-165568-         907 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Empty concatmer vector in which 2 gRNAs can be inserted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 907)
  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 907)
  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: 2 gRNA concatmer
FEATURES             Location/Qualifiers
     source          1..907
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        14..262
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        279..354
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             361..384
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        407..655
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        674..749
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             755..781
                     /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             755..781
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(833..856)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ggtcgggcag gaagagggcc tatttcccat gattccttca tatttgcata tacgatacaa
       61 ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa
      121 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      181 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      241 cttgtggaaa ggacgaaaca ccgggtcttc gagaagacgt tttagagcta gaaatagcaa
      301 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
      361 gactacaagg acgacgatga caaggatctc caaggtcggg caggaagagg gcctatttcc
      421 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa
      481 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
      541 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
      601 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggagt
      661 cttcgagaag acagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
      721 aacttgaaaa agtggcaccg agtcggtgct tttttacccc tacgacgtgc ccgactacgc
      781 cggatccgaa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc
      841 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc
      901 cacatcc
//
