LOCUS       sequence-86619-e         868 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses the S. pyogenes sgRNA from the 7SK promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 868)
  AUTHORS   Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA
  TITLE     Multiplex CRISPR/Cas9-based genome engineering from a single 
            lentiviral vector.
  JOURNAL   Nucleic Acids Res. 2014 Aug 13. pii: gku749.
  PUBMED    25122746
REFERENCE   2  (bases 1 to 868)
  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. 2014 Aug 13. pii: gku749."
COMMENT     Sequence Label: ph7SK-gRNA
FEATURES             Location/Qualifiers
     source          1..868
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        321..396
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(452..472)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(452..470)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(491..507)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(491..507)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(504..526)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    515..531
                     /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(539..545,546..563,564..569))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    584..605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(722..739)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gactcactat agggcgaatt gggtaccttt aattctagta ctatgcatcg tctcattgtc
       61 tgcagtattt agcatgcccc acccatctgc aaggcattct ggatagtgtc aaaacagccg
      121 gaaatcaagt ccgtttatct caaactttag cattttggga ataaatgata tttgctatgc
      181 tggttaaatt agattttagt taaatttcct gctgaagctc tagtacgata agcaacttga
      241 cctaagtgta aagttgagac ttccttcagg tttatatagc ttgtgcgccg cttgggtacc
      301 tcgggtcttc gagaagacct gttttagagc tagaaatagc aagttaaaat aaggctagtc
      361 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttctcggac tgatgacctg
      421 agacgccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
      481 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      541 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      601 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      661 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      721 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      781 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      841 caaaaggcca gcaaaaggcc aggaaccg
//
