LOCUS       sequence-86612-e         885 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses the S. pyogenes sgRNA from the H1 promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 885)
  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 885)
  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: phH1-gRNA
FEATURES             Location/Qualifiers
     source          1..885
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        31..245
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     166..185
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        272..347
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(392..412)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(392..410)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(431..447)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(431..447)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(444..466)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    455..471
                     /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(479..485,486..503,504..509))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    524..545
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(662..679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tatagggcga attgggtacc cgtctcacct gaacgctgac gtcatcaacc cgctccaagg
       61 aatcgcgggc ccagtgtcac taggcgggaa cacccagcgc gcgtgcgccc tggcaggaag
      121 atggctgtga gggacagggg agtggcgccc tgcaatattt gcatgtcgct atgtgttctg
      181 ggaaatcacc ataaacgtga aatgtctttg gatttgggaa tcttataagt tctgtatgag
      241 accactcttt cccaggtctt cgagaagacc tgttttagag ctagaaatag caagttaaaa
      301 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttggtctg
      361 agacgccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
      421 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      481 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      541 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      601 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      661 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      721 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      781 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      841 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtc
//
