LOCUS       sequence-108430-         716 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA with 2x MS2 (wt+f6) for yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 716)
  AUTHORS   Zalatan JG, Lee ME, Almeida R, Gilbert LA, Whitehead EH, La Russa M,
            Tsai JC, Weissman JS, Dueber JE, Qi LS, Lim WA
  TITLE     Engineering Complex Synthetic Transcriptional Programs with CRISPR 
            RNA Scaffolds.
  JOURNAL   Cell. 2014 Dec 18. pii: S0092-8674(14)01570-0. doi: 
            10.1016/j.cell.2014.11.052.
  PUBMED    25533786
REFERENCE   2  (bases 1 to 716)
  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: "Cell. 2014 
            Dec 18. pii: S0092-8674(14)01570-0. doi: 
            10.1016/j.cell.2014.11.052."
COMMENT     Sequence Label: pJZC588
FEATURES             Location/Qualifiers
     source          1..716
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        17..285
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        306..381
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        388..406
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      438..457
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        complement(486..504)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(525..541)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(525..541)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(538..560)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    549..565
                     /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(573..579,580..597,598..603))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    618..639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 agctcactcg gaattctctt tgaaaagata atgtatgatt atgctttcac tcatatttat
       61 acagaaactt gatgttttct ttcgagtata tacaaggtga ttacatgtac gtttgaagta
      121 caactctaga ttttgtagtg ccctcttggg ctagcggtaa aggtgcgcat tttttcacac
      181 cctacaatgt tctgttcaaa agattttggt caaacgctgt agaagtgaaa gttggtgcgc
      241 atgtttcggc gttcgaaact tctccgcagt gaaagataaa tgatcacttt tctctatcac
      301 tgatagtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
      361 aaagtggcac cgagtcggtg cgggagcaca tgaggatcac ccatgtgcga ctcccacagt
      421 cactggggag tcttcccttt ttttgttttt tatgtctctg cagagttcgg taccagcttt
      481 tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct
      541 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      601 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
      661 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgc
//
