LOCUS       sequence-148116-         918 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of customizable guide RNA (gRNA) for bacterial gene 
            knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 918)
  AUTHORS   Qi LS, Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim 
            WA
  TITLE     Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific 
            Control of Gene Expression.
  JOURNAL   Cell. 2013 Feb 28;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022.
  PUBMED    23452860
REFERENCE   2  (bases 1 to 918)
  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.cell.2013.02"; journalName: "Cell"; date: "2013-02-28- 
            28"; volume: "152"; issue: "5"; pages: "1173-83"
COMMENT     Sequence Label: pgRNA-bacteria
FEATURES             Location/Qualifiers
     source          1..918
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(191..295)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     363..381
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(385..428)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        446..480
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        501..576
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             597..626
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             642..659
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(715..732)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
       61 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
      121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
      181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
      241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
      301 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
      361 aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct ttcgctaagg
      421 atgatttctg gaattctaaa gatctttgac agctagctca gtcctaggta taatactagt
      481 aactttcagt ttagcggtct gttttagagc tagaaatagc aagttaaaat aaggctagtc
      541 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttgaagctt gggcccgaac
      601 aaaaactcat ctcagaagag gatctgaata gcgccgtcga ccatcatcat catcatcatt
      661 gagtttaaac ggtctccagc ttggctgttt tggcggatga gagaagattt tcagcctgat
      721 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
      781 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
      841 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
      901 ctcagtcgaa agactggg
//
