LOCUS       sequence-99973-e         803 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 803)
  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 803)
  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..803
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(169..188)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(330..347)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(330..346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(339..361)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(555..574)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     674..696
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(734..752)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ccgactcggt gccacttttt caagttgata acggactagc cttattttaa cttgctattt
       61 ctagctctaa aacagagacc actagtggca caggtctcag gggatctgtg gtcctcatac
      121 agaacttata agattcccaa atccaaagac atttcacgtt tatggtgatt tcccagaaca
      181 catagcgaca tgcaaatatt gcagggcgcc actcccctgt ccctcacagc catcttcctg
      241 ccagggcgca cgcgcgctgg gtgttcccgc ctagtgacac tgggcccgcg attccttgga
      301 gcgggttgat gacgtcagcg ttcgaattca ctggccgtcg ttttacaacg tcgtgactgg
      361 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
      421 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
      481 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
      541 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
      601 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
      661 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
      721 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
      781 gtttcttaga cgtcaggtgg cac
//
