LOCUS       sequence-124641-         976 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Transient expression in mammalian cells of an ""INT"" 
            construct_bearing P4_P6 (internally appended with three PP7 
            stem-loops), targeting the GLuc reporter. U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 976)
  AUTHORS   Shechner DM, Hacisuleyman E, Younger ST, Rinn JL
  TITLE     Multiplexable, locus-specific targeting of long RNAs with 
            CRISPR-Display.
  JOURNAL   Nat Methods. 2015 Jul;12(7):664-70. doi: 10.1038/nmeth.3433. Epub 
            2015 Jun 1.
  PUBMED    26030444
REFERENCE   2  (bases 1 to 976)
  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: 
            "10.1038/nmeth.3433"; journalName: "Nat Methods"; date: "2015-07"; 
            volume: "12"; issue: "7"; pages: "664-70"
COMMENT     Sequence Label: pU6_(GLuc)_INT[P4-P6(3xPP7)]
FEATURES             Location/Qualifiers
     source          1..976
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(473..489)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(473..489)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(486..508)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    497..513
                     /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)."
     promoter        complement(join(521..527,528..545,546..551))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    566..587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(704..721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(955..974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggctttatat atcttgtgga aaggacgaaa caccgatcta gatacgactc actatgttta
       61 agagctatgc tgcgaatacg agtctggaat tgcgggaaag gggtcaacag ccgttcagta
      121 ccaagtctca ggggaaactt tgagatggcc ttgcaaaggg tatggtaata agctgacgga
      181 catggtccta acacgcagcc aagtcctaag tcaacagtct ggagcagacg atatggcgtc
      241 gctcctctcc acgagagcat atgggctccg tggtctccag cagacgatat ggcgtcgctg
      301 gtctctgttg atatggatgc agttcatctc tcgtattcgc agcatagcaa gtttaaataa
      361 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tcttaattaa
      421 gtctagagtc gactgtttaa acctgcaggc atgcaagctt ggcgtaatca tggtcatagc
      481 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      541 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      601 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      661 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      721 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      781 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      841 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      901 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      961 accaggcgtt tccccc
//
