LOCUS       sequence-99972-e         818 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPR/Cas9 system gRNA cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 818)
  AUTHORS   Zheng Q, Cai X, Tan MH, Schaffert S, Arnold CP, Gong X, Chen CZ, 
            Huang S
  TITLE     Precise gene deletion and replacement using the CRISPR/Cas9 system 
            in human cells.
  JOURNAL   Biotechniques. 2014 Sep 1;57(3):115-24. doi: 10.2144/000114196. 
            eCollection 2014.
  PUBMED    25209046
REFERENCE   2  (bases 1 to 818)
  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"; doi: "10.2144/000114196";
            journalName: "Biotechniques"; date: "2014-09-1- 1"; volume: "57"; 
            issue: "3"; pages: "115-24"
COMMENT     Sequence Label: pUC-H1-gRNA
FEATURES             Location/Qualifiers
     source          1..818
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     231..248
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    365..386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(401..406,407..424,425..431)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    439..455
                     /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)."
     primer_bind     444..466
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     463..479
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     463..479
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        complement(512..587)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(683..702)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
ORIGIN
        1 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
       61 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
      121 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
      181 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
      241 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
      301 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
      361 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
      421 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
      481 atgattacgc caagcttcac gcgtaaaaaa agcaccgact cggtgccact ttttcaagtt
      541 gataacggac tagccttatt ttaacttgct atttctagct ctaaaacaga gaccactagt
      601 ggcacaggtc tcaggggatc tgtggtcctc atacagaact tataagattc ccaaatccaa
      661 agacatttca cgtttatggt gatttcccag aacacatagc gacatgcaaa tattgcaggg
      721 cgccactccc ctgtccctca cagccatctt cctgccaggg cgcacgcgcg ctgggtgttc
      781 ccgcctagtg acactgggcc cgcgattcct tggagcgg
//
