LOCUS       sequence-97844-e         960 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  gRNA expression by the H1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 960)
  AUTHORS   Ranganathan V, Wahlin K, Maruotti J, Zack DJ
  TITLE     Expansion of the CRISPR-Cas9 genome targeting space through the use 
            of H1 promoter-expressed guide RNAs.
  JOURNAL   Nat Commun. 2014 Aug 8;5:4516. doi: 10.1038/ncomms5516.
  PUBMED    25105359
REFERENCE   2  (bases 1 to 960)
  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: "Nat 
            Commun."; date: "2014-08-8"; volume: "5:4516. doi"; pages: " 
            10.1038/ncomms5516"
COMMENT     Sequence Label: pH1v1
FEATURES             Location/Qualifiers
     source          1..960
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        54..129
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(141..165)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(212..228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(212..228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(225..247)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    236..252
                     /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(260..266,267..284,285..290))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    305..326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(443..460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(694..713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtctttggat ttgggaatct tataagttct gtatgagacc actttttccc taggttttag
       61 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
      121 agtcggtgct ttttttctag acccagcttt cttgtacaaa gttggcatta ctagagtcga
      181 cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
      241 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
      301 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
      361 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
      421 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      481 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      541 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      781 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
//
