LOCUS       sequence-131008-         718 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses palindromic sgRNA for use in self-cloning CRISPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 718)
  AUTHORS   Arbab M, Srinivasan S, Hashimoto T, Geijsen N, Sherwood RI
  TITLE     Cloning-free CRISPR.
  JOURNAL   Stem Cell Reports. 2015 Oct 27. pii: S2213-6711(15)00284-2. doi: 
            10.1016/j.stemcr.2015.09.022.
  PUBMED    26527385
REFERENCE   2  (bases 1 to 718)
  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: "Stem Cell 
            Reports. 2015 Oct 27. pii: S2213-6711(15)00284-2. doi: 
            10.1016/j.stemcr.2015.09.022."
COMMENT     Sequence Label: sgPal7 (p2Tol-U6-sgPal7-HygR)
FEATURES             Location/Qualifiers
     source          1..718
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(45..293)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(103..122)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(273..293)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     protein_bind    311..344
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    382..415
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      421..430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(455..617,618..629)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(659..678)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(660..678)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(688..705)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(688..704)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgtttccagc atagctctta aacgcttagt actagtacta agccggtgtt tcgtcctttc
       61 cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag catatgatag
      121 tccattttaa aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt
      181 cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattccaat tatctctcta
      241 acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc ctcacatgcg
      301 cgccggtacc ataacttcgt ataaagtatc ctatacgaag ttatggccgg ccaagggcaa
      361 ttccaccaca ctggactatc gataacttcg tataatgtat gctatacgaa gttatcgaca
      421 ccatggtggc ctccagatcc ttcgagatcg atctttcttg cttttacttt tacttcctta
      481 atactcaagt acaattttaa tggagtactt ttttactttt actcaagtaa gattctagcc
      541 agatactttt acttttaatt gagtaaaatt ttccctaagt acttgtactt tcacttgagt
      601 aaaatttttg agtacttttt acacctctgt caagaaccat atgccggtac ccaattcgcc
      661 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactgg
//
