LOCUS       sequence-135016-         939 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 (2 Gs at 
            initiation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 939)
  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 939)
  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: pU6T7
FEATURES             Location/Qualifiers
     source          1..939
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     44..63
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(194..216)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     316..335
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     529..551
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     611..631
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     782..801
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        843..861
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccatgttgga atttaatcgc ggcctcgacg tttcccgttg aatatggctc ataacacccc
       61 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
      121 gtgcaatgta acatcagaga ttttgagaca cgggccagag ctgcatcgcg cgtttcggtg
      181 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
      241 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
      301 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
      361 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
      421 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      481 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      541 gttgtaaaac gacggccaga gaattcgagc tcggtacctc gcgaatacat ctagaaaggt
      601 cgggcaggaa gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
      661 tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
      721 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
      781 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
      841 gctaatacga ctcactatag gagagaccga gagagggtct cagttttaga gctagaaata
      901 gcaagttaaa ataaggctag tccgttatca acttgaaaa
//
