LOCUS       sequence-107470-         787 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PCR template to construct polycistronic tRNA-gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 787)
  AUTHORS   Xie K, Minkenberg B, Yang Y
  TITLE     Boosting CRISPR/Cas9 multiplex editing capability with the 
            endogenous tRNA-processing system.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 
            10.1073/pnas.1420294112. Epub 2015 Mar 2.
  PUBMED    25733849
REFERENCE   2  (bases 1 to 787)
  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: 
            "10.1073/pnas.1420294112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-03-17- 17"; volume: "112"; issue: "11"; pages: "3570-5"
COMMENT     Sequence Label: pGTR
FEATURES             Location/Qualifiers
     source          1..787
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     186..208
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     200..217
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     201..217
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     224..243
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        224..242
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      273..282
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      277..286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_RNA        complement(385..460)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(557..575)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(558..575)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(593..609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(593..609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(606..628)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    617..633
                     /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(641..647,648..665,666..671))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    686..707
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
       61 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      121 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      181 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata
      241 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
      301 tctgccatgc accagccggg aatcgaaccc gggtctgtac cgtggcaggg tactattcta
      361 ccactagacc actggtgctt tgttgcaccg actcggtgcc actttttcaa gttgataacg
      421 gactagcctt attttaactt gctatttcta gctctaaaac cgagacctcg gtctccaatc
      481 actagtgaat tcgcggccgc ctgcaggtcg accatatggg agagctccca acgcgttgga
      541 tgcatagctt gagtattcta tagtgtcacc taaatagctt ggcgtaatca tggtcatagc
      601 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      661 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      781 gcgcggg
//
