LOCUS       sequence-162993-        1222 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Empty concatmer vector in which 3 gRNAs can be inserted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1222)
  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 1222)
  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: 3 gRNA concatemer
FEATURES             Location/Qualifiers
     source          1..1222
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        24..272
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        289..364
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             371..394
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        417..665
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        684..759
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             765..791
                     /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             765..791
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        815..1063
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        1084..1159
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 agatctccaa ggtcgggcag gaagagggcc tatttcccat gattccttca tatttgcata
       61 tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat
      121 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
      181 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
      241 ctttatatat cttgtggaaa ggacgaaaca ccgggtcttc gagaagacgt tttagagcta
      301 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
      361 gtgctttttt gactacaagg acgacgatga caaggatctc caaggtcggg caggaagagg
      421 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
      481 ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
      541 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
      601 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
      661 acaccggagt cttcgagaag acagttttag agctagaaat agcaagttaa aataaggcta
      721 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttacccc tacgacgtgc
      781 ccgactacgc cggatctcca aggtcgggca ggaagagggc ctatttccca tgattccttc
      841 atatttgcat atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa
      901 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
      961 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
     1021 cgatttcttg gctttatata tcttgtggaa aggacgaaac accggaagtc ttcgagaaga
     1081 caagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     1141 agtggcaccg agtcggtgct tttttatggc atcaatgcag aagctgatct cagaggagga
     1201 cctgggatcc gaattcgtcg ac
//
