LOCUS       sequence-124353-         866 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 866)
  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 866)
  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..866
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     216..233
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    350..371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(386..391,392..409,410..416)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    424..440
                     /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     429..451
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     448..464
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     448..464
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        482..500
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     482..499
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    579..603
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(615..690)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(769..788)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
       61 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      121 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      181 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      241 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      301 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      361 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      421 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      481 tatttaggtg acactataga atactcaagc tatgcatcca acgcgttggg agctctccca
      541 tatggtcgac ctgcaggcgg ccgcactagt gattaatgcc aactttgtac aagaaagctg
      601 ggtctagaaa aaaagcaccg actcggtgcc actttttcaa gttgataacg gactagcctt
      661 attttaactt gctatttcta gctctaaaac gactgtaagt gaattacttc ggtgtttcgt
      721 cctttccaca agatatataa agccaagaaa tcgaaatact ttcaagttac ggtaagcata
      781 tgatagtcca ttttaaaaca taattttaaa actgcaaact acccaagaaa ttattacttt
      841 ctacgtcacg tattttgtac taatat
//
