LOCUS       sequence-86614-e         887 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses the S. pyogenes sgRNA from the mouse U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 887)
  AUTHORS   Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA
  TITLE     Multiplex CRISPR/Cas9-based genome engineering from a single 
            lentiviral vector.
  JOURNAL   Nucleic Acids Res. 2014 Aug 13. pii: gku749.
  PUBMED    25122746
REFERENCE   2  (bases 1 to 887)
  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: "Nucleic 
            Acids Res. 2014 Aug 13. pii: gku749."
COMMENT     Sequence Label: pmU6-gRNA
FEATURES             Location/Qualifiers
     source          1..887
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        36..349
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     268..288
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        367..442
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(487..507)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(487..505)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(526..542)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(526..542)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(539..561)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    550..566
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(574..580,581..598,599..604))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    619..640
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(757..774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcactatagg gcgaattggg tacccgtctc agatagatcc gacgccgcca tctctaggcc
       61 cgcgccggcc ccctcgcaca gacttgtggg agaagctcgg ctactcccct gccccggtta
      121 atttgcatat aatatttcct agtaactata gaggcttaat gtgcgataaa agacagataa
      181 tctgttcttt ttaatactag ctacatttta catgataggc ttggatttct ataagagata
      241 caaatactaa attattattt taaaaaacag cacaaaagga aactcaccct aactgtaaag
      301 taattgtgtg ttttgagact ataaatatcc cttggagaaa agccttgttt gtcttcgaga
      361 agacctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      421 aaaagtggca ccgagtcggt gctttttttg tactgagacg ccgcggtgga gctccagctt
      481 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
      541 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      601 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      661 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
      721 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
      781 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      841 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagc
//
