LOCUS       sequence-124471-        1061 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Transient expression of the ""TOP1"" construct, targeting the GLuc 
            reporter, in mammalian cells. CMV/3' Box expression backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1061)
  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 1061)
  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: pCMV/3' Box_(GLuc)_TOP1
FEATURES             Location/Qualifiers
     source          1..1061
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        64..139
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(572..588)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(572..588)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(585..607)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    596..612
                     /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(620..626,627..644,645..650))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    665..686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(803..820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcagagctct ctggctaact agagaaccca ctgcttactg gcgatctaga tacgactcac
       61 tatgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
      121 agtggcaccg agtcggtgct cttcaacatc cctcctctaa acccggaatt gcgggaaagg
      181 ggtcaacagc cgttcagtac caagtctcag gggaaacttt gagatggcct tgcaaagggt
      241 atggtaataa gctgacggac atggtcctaa cacgcagcca agtcctaagt caacagtttt
      301 tggagcagac gatatggcgt cgctcctttt tccagcagag catatgggct ccgtggtttt
      361 tccagcagac gatatggcgt cgctggtttt tggagcagag catatgggct cgctcctttt
      421 tgcagcagac gatatggcgt cgctgctttt tctgttgata tggatgcagt tcaactttct
      481 ggagtttcaa aagtagactg tacgctaagg gtcatatctt tttttgtttg gtttgtgtct
      541 tggttggcgt cttaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
      601 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
      661 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
      721 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
      781 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      841 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      901 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      961 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1021 caagtcagag gtggcgaaac ccgacaggac tataaagata c
//
