LOCUS       sequence-131004-         712 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 712)
  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 712)
  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: sgPal1 (p2Tol-U6-sgPal1-HygR)
FEATURES             Location/Qualifiers
     source          1..712
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(39..287)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(97..116)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(267..287)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     protein_bind    305..338
                     /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    376..409
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      415..424
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(449..611,612..623)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(653..672)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(654..672)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(682..699)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(682..698)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cagcatagct cttaaacctc tgtgactagt cacagagcgg tgtttcgtcc tttccacaag
       61 atatataaag ccaagaaatc gaaatacttt caagttacgg taagcatatg atagtccatt
      121 ttaaaacata attttaaaac tgcaaactac ccaagaaatt attactttct acgtcacgta
      181 ttttgtacta atatctttgt gtttacagtc aaattaattc caattatctc tctaacagcc
      241 ttgtatcgta tatgcaaata tgaaggaatc atgggaaata ggccctcaca tgcgcgccgg
      301 taccataact tcgtataaag tatcctatac gaagttatgg ccggccaagg gcaattccac
      361 cacactggac tatcgataac ttcgtataat gtatgctata cgaagttatc gacaccatgg
      421 tggcctccag atccttcgag atcgatcttt cttgctttta cttttacttc cttaatactc
      481 aagtacaatt ttaatggagt acttttttac ttttactcaa gtaagattct agccagatac
      541 ttttactttt aattgagtaa aattttccct aagtacttgt actttcactt gagtaaaatt
      601 tttgagtact ttttacacct ctgtcaagaa ccatatgccg gtacccaatt cgccctatag
      661 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gg
//
