LOCUS       sequence-135018-         959 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses gRNA from hybrid human U6/T7 promoter for both cellular 
            expression and in vitro transcription from same promoter (1 G at 
            initiation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 959)
  AUTHORS   Romanienko PJ, Giacalone J, Ingenito J, Wang Y, Isaka M, Johnson T, 
            You Y, Mark WH
  TITLE     A Vector with a Single Promoter for In Vitro Transcription and 
            Mammalian Cell Expression of CRISPR gRNAs.
  JOURNAL   PLoS One. 2016 Feb 5;11(2):e0148362. doi: 
            10.1371/journal.pone.0148362. eCollection 2016.
  PUBMED    26849369
REFERENCE   2  (bases 1 to 959)
  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: "PLoS One.";
            date: "2016-02-5"; volume: "11(2):e0148362. doi"; pages: " 
            10.1371/journal.pone.0148362. eCollection 2016"
COMMENT     Sequence Label: pU6T7G
FEATURES             Location/Qualifiers
     source          1..959
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     41..60
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(191..213)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     313..332
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     526..548
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     608..628
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     779..798
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     840..859
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        840..858
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        879..954
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tgttggaatt taatcgcggc ctcgacgttt cccgttgaat atggctcata acaccccttg
       61 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
      121 caatgtaaca tcagagattt tgagacacgg gccagagctg catcgcgcgt ttcggtgatg
      181 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
      241 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
      301 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
      361 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
      421 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
      481 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
      541 gtaaaacgac ggccagagaa ttcgagctcg gtacctcgcg aatacatcta gaaaggtcgg
      601 gcaggaagag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt
      661 tagagagata attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg
      721 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
      781 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgct
      841 aatacgactc actataggga gaccgagaga gggtctcagt tttagagcta gaaatagcaa
      901 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgcttttt
//
