LOCUS       sequence-148117-         911 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 911)
  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 911)
  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..911
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     210..227
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(280..326)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     479..496
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(552..569)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(585..614)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_RNA        complement(635..710)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(731..765)
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     terminator      783..826
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(830..848)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagcctg tggaaaaacg
       61 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      121 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      181 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcctgcagtc
      241 tagactcgag taaggatcca gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
      301 ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg catggggaga
      361 ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgg ggtcaggtgg
      421 gaccaccgcg ctactgccgc caggcaaatt ctgttttatc agaccgcttc tgcgttctga
      481 tttaatctgt atcaggctga aaatcttctc tcatccgcca aaacagccaa gctggagacc
      541 gtttaaactc aatgatgatg atgatgatgg tcgacggcgc tattcagatc ctcttctgag
      601 atgagttttt gttcgggccc aagcttcaaa aaaagcaccg actcggtgcc actttttcaa
      661 gttgataacg gactagcctt attttaactt gctatttcta gctctaaaac agaccgctaa
      721 actgaaagtt actagtatta tacctaggac tgagctagct gtcaaagatc tttagaattc
      781 cagaaatcat ccttagcgaa agctaaggat tttttttatc tgaaattctg cctcgtgata
      841 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
      901 tttcggggaa a
//
