LOCUS       sequence-124638-        1009 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Transient expression in mammalian cells of an ""INT"" 
            construct_bearing the ""Bunch of Baby Spinach"" (or ""BoBS"") 
            construct, targeting the GLuc reporter. U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1009)
  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 1009)
  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[BoBS]
FEATURES             Location/Qualifiers
     source          1..1009
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(362..378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(362..378)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(375..397)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    386..402
                     /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(410..416,417..434,435..440))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    455..476
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(593..610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(844..863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atttcttggc tttatatatc ttgtggaaag gacgaaacac cgatctagat acgactcact
       61 atgtttaaga gctatgctgc gaatacgaga aggacgggtc cggacgcaag gacgggtccg
      121 accgaaagga cgggtccaat ggtggaaaca ccattgttga gtagagtgtg agtcggtcgt
      181 tgagtagagt gtgaggcgtc cgttgagtag agtgtgagct cgtattcgca gcatagcaag
      241 tttaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt
      301 cttaattaag tctagagtcg actgtttaaa cctgcaggca tgcaagcttg gcgtaatcat
      361 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      421 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      481 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
      541 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
      601 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      661 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
      721 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      781 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
      841 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
      901 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
      961 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagct
//
