LOCUS       sequence-98841-e         592 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  crRNA expression plasmid on pRS423 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 592)
  AUTHORS   Bao Z, Xiao H, Liang J, Zhang L, Xiong X, Sun N, Si T, Zhao H
  TITLE     Homology-Integrated CRISPR-Cas (HI-CRISPR) System for One-Step 
            Multigene Disruption in Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2014 Sep 19.
  PUBMED    25207793
REFERENCE   2  (bases 1 to 592)
  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: "ACS Synth 
            Biol. 2014 Sep 19."
COMMENT     Sequence Label: pCR
FEATURES             Location/Qualifiers
     source          1..592
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     11..30
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        11..29
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(77..97)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(77..95)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(116..132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    140..156
                     /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(164..170,171..188,189..194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    209..230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      303..322
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     primer_bind     complement(360..376)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    384..400
                     /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(408..414,415..432,433..438))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    453..474
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt
       61 cctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
      121 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
      181 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
      241 gctcactgcc cgctttccac cggtggtctc tagagctatg ctgttttgaa tggtcccaaa
      301 actttttttg ttttttatgt ctgagctccc tgcaggcatg caagcttggc gtaatcatgg
      361 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
      421 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
      481 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
      541 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ct
//
