LOCUS       sequence-125086-        1110 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  gRNA targeting human HPRT gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1110)
  AUTHORS   Liao S, Tammaro M, Yan H
  TITLE     Enriching CRISPR-Cas9 targeted cells by co-targeting the HPRT gene.
  JOURNAL   Nucleic Acids Res. 2015 Jun 29. pii: gkv675.
  PUBMED    26130722
REFERENCE   2  (bases 1 to 1110)
  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. 2015 Jun 29. pii: gkv675."
COMMENT     Sequence Label: HPRT2b-pGEM
FEATURES             Location/Qualifiers
     source          1..1110
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     133..152
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        231..306
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(318..342)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(421..439)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(422..439)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(457..473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(457..473)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(470..492)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    481..497
                     /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(505..511,512..529,530..535))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    550..571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(688..705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(939..958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
       61 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      121 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      181 ttatatatct tgtggaaagg acgaaacacc gaagtaattc acttacagtc gttttagagc
      241 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      301 cggtgctttt tttctagacc cagctttctt gtacaaagtt ggcattaatc actagtgcgg
      361 ccgcctgcag gtcgaccata tgggagagct cccaacgcgt tggatgcata gcttgagtat
      421 tctatagtgt cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      481 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      541 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      601 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      661 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      721 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      781 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      841 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      901 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      961 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1021 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1081 ggtgtaggtc gttcgctcca agctgggctg
//
