LOCUS       sequence-105885-         880 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 880)
  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 880)
  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..880
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        complement(100..175)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(272..290)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(273..290)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(308..324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(308..324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(321..343)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    332..348
                     /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(356..362,363..380,381..386))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    401..422
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(539..556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(790..809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgcgggaat tcgattctgc catgcaccag ccgggaatcg aacccgggtc tgtaccgtgg
       61 cagggtacta ttctaccact agaccactgg tgctttgttg caccgactcg gtgccacttt
      121 ttcaagttga taacggacta gccttatttt aacttgctat ttctagctct aaaaccgaga
      181 cctcggtctc caatcactag tgaattcgcg gccgcctgca ggtcgaccat atgggagagc
      241 tcccaacgcg ttggatgcat agcttgagta ttctatagtg tcacctaaat agcttggcgt
      301 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      361 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
      421 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      481 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      541 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      601 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      661 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      721 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      781 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      841 cgaccctgcc gcttaccgga tacctgtccg cctttctccc
//
