LOCUS       sequence-112389-        1036 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cloning and expression of Streptococcus pyogenes guide RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1036)
  AUTHORS   Ding Q, Regan SN, Xia Y, Oostrom LA, Cowan CA, Musunuru K
  TITLE     Enhanced efficiency of human pluripotent stem cell genome editing 
            through replacing TALENs with CRISPRs.
  JOURNAL   Cell Stem Cell. 2013 Apr 4;12(4):393-4. doi: 
            10.1016/j.stem.2013.03.006.
  PUBMED    23561441
REFERENCE   2  (bases 1 to 1036)
  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: 
            "10.1016/j.stem.2013.03"; journalName: "Cell Stem Cell"; date: 
            "2013-04-4- 4"; volume: "12"; issue: "4"; pages: "393-4"
COMMENT     Sequence Label: pGuide
FEATURES             Location/Qualifiers
     source          1..1036
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     223..240
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    357..378
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(393..398,399..416,417..423)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    431..447
                     /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     436..458
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     455..471
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     455..471
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        489..507
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     489..506
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    595..619
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(631..706)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(725..973)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(783..802)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(953..973)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
       61 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
      121 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
      181 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
      241 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
      301 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      361 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      421 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
      481 gccaagctat ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag
      541 ctcggatcca ctagtaacgg ccgccagtgt gctggaattc gcccttcatt aatgccaact
      601 ttgtacaaga aagctgggtc tagaaaaaaa gcaccgactc ggtgccactt tttcaagttg
      661 ataacggact agccttattt taacttgcta tttctagctc taaaacaggt cttctcgaag
      721 acccggtgtt tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa tactttcaag
      781 ttacggtaag catatgatag tccattttaa aacataattt taaaactgca aactacccaa
      841 gaaattatta ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat
      901 taattctaat tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg
      961 gaaataggcc ctcttcctgc ccgaccttgg taccggatcc agtcgactga attggttcct
     1021 ttaaagcctg cttttt
//
