LOCUS       sequence-165569-         913 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 913)
  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 913)
  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..913
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(35..54)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     428..451
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             complement(503..529)
                     /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             complement(503..529)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_RNA        complement(535..610)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(629..877)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(687..706)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(857..877)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 atgaggaaga ggagaacagc gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc
       61 ctccggagga ccttcgggcg cccgccccgc ccctgagccc gcccctgagc ccgcccccgg
      121 acccacccct tcccagcctc tgagcccaga aagcgaagga gcaaagctgc tattggccgc
      181 tgccccaaag gcctacccgc ttccattgct cagcggtgct gtccatctgc acgagactag
      241 tgagacgtgc tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga
      301 acttcctgac taggggagga gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg
      361 gttggcgcct accggtggat gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc
      421 aagtgcccag cggggctgct aaagcgcatg ctccagactg ccttgggaaa agcgcctccc
      481 ctacccggta gaattcggat ccggcgtagt cgggcacgtc gtaggggtaa aaaagcaccg
      541 actcggtgcc actttttcaa gttgataacg gactagcctt attttaactt gctatttcta
      601 gctctaaaac tgtcttctcg aagactccgg tgtttcgtcc tttccacaag atatataaag
      661 ccaagaaatc gaaatacttt caagttacgg taagcatatg atagtccatt ttaaaacata
      721 attttaaaac tgcaaactac ccaagaaatt attactttct acgtcacgta ttttgtacta
      781 atatctttgt gtttacagtc aaattaattc taattatctc tctaacagcc ttgtatcgta
      841 tatgcaaata tgaaggaatc atgggaaata ggccctcttc ctgcccgacc ttggagatcc
      901 ttgtcatcgt cgt
//
