LOCUS       sequence-131011-         712 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Base plasmid for sgRNA cloning with Hygromycin resistance and Tol2 
            transposon arms.
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: sgBbsI (p2Tol-U6-2xBbsI-sgRNA-HygR)
FEATURES             Location/Qualifiers
     source          1..712
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(36..284)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(94..113)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(264..284)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     protein_bind    302..335
                     /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    373..406
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      412..421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(446..608,609..620)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(650..669)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(651..669)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(679..696)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(679..695)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(688..710)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cagcatagct cttaaacagg tcttctcgaa gacccggtgt ttcgtccttt ccacaagata
       61 tataaagcca agaaatcgaa atactttcaa gttacggtaa gcatatgata gtccatttta
      121 aaacataatt ttaaaactgc aaactaccca agaaattatt actttctacg tcacgtattt
      181 tgtactaata tctttgtgtt tacagtcaaa ttaattccaa ttatctctct aacagccttg
      241 tatcgtatat gcaaatatga aggaatcatg ggaaataggc cctcacatgc gcgccggtac
      301 cataacttcg tataaagtat cctatacgaa gttatggccg gccaagggca attccaccac
      361 actggactat cgataacttc gtataatgta tgctatacga agttatcgac accatggtgg
      421 cctccagatc cttcgagatc gatctttctt gcttttactt ttacttcctt aatactcaag
      481 tacaatttta atggagtact tttttacttt tactcaagta agattctagc cagatacttt
      541 tacttttaat tgagtaaaat tttccctaag tacttgtact ttcacttgag taaaattttt
      601 gagtactttt tacacctctg tcaagaacca tatgccggta cccaattcgc cctatagtga
      661 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aa
//
