LOCUS       sequence-135017-        1002 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 1002)
  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 1002)
  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..1002
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     215..232
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    349..370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(385..390,391..408,409..415)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    423..439
                     /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)."
     primer_bind     428..450
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     447..463
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     447..463
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        complement(538..613)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(633..652)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(634..652)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(694..713)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(864..884)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(944..966)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
       61 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      121 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      181 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      241 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
      301 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
      361 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      421 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acaccaagct
      481 tgcatgcagg cctctgcagt cgacgggccc gggatccgat atctagattt aaaaaaagca
      541 ccgactcggt gccacttttt caagttgata acggactagc cttattttaa cttgctattt
      601 ctagctctaa aactgagacc ctctctcggt ctccctatag tgagtcgtat tagcaagata
      661 tataaagcca agaaatcgaa atactttcaa gttacggtaa gcatatgata gtccatttta
      721 aaacataatt ttaaaactgc aaactaccca agaaattatt actttctacg tcacgtattt
      781 tgtactaata tctttgtgtt tacagtcaaa ttaattctaa ttatctctct aacagccttg
      841 tatcgtatat gcaaatatga aggaatcatg ggaaataggc cctcttcctg cccgaccttt
      901 ctagatgtat tcgcgaggta ccgagctcga attctctggc cgtcgtttta caacgtcgtg
      961 actgggaaaa ccctggcgtt acccaactta atcgccttgc ag
//
