LOCUS       sequence-72728-e         992 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian U6-driven sgRNA (STm1) targeting GTCCCCTCCACCCCACAGTG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 992)
  AUTHORS   Esvelt KM, Mali P, Braff JL, Moosburner M, Yaung SJ, Church GM
  TITLE     Orthogonal Cas9 proteins for RNA-guided gene regulation and editing.
  JOURNAL   Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.
  PUBMED    24076762
REFERENCE   2  (bases 1 to 992)
  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: "Nat 
            Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681."
COMMENT     Sequence Label: M-ST1-sgRNA
FEATURES             Location/Qualifiers
     source          1..992
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        57..181
                     /label=St1 gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus 
                     thermophilus CRISPR/Cas9 system (Esvelt et al., 2013)"
     protein_bind    complement(193..217)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(262..278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(262..278)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(275..297)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    286..302
                     /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(310..316,317..334,335..340))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    355..376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(493..510)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(744..763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttggcttta tatatcttgt ggaaggacga aacaccgtcc cctccacccc acagtggttt
       61 ttgtactctc aagatttaag taactgtaca acgaaactta cacagttact taaatcttgc
      121 agaagctaca aagataaggc ttcatgccga aatcaacacc ctgtcatttt atggcagggt
      181 gtttttttct agacccagct ttcttgtaca aagttggcat tagttccatc ccaatggcgc
      241 gccgagcttg gctcgagcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
      301 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
      361 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
      421 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
      481 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      541 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      601 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      841 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatcccg
      961 gtaactatcg tcttgagtcc aacccggtaa ga
//
