LOCUS       sequence-98908-e         686 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid encoding iCas9 and tracrRNA on pRS415 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 686)
  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 686)
  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: pCT
FEATURES             Location/Qualifiers
     source          1..686
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     85..102
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    219..240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(255..260,261..278,279..285)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    293..309
                     /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)."
     primer_bind     298..320
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     317..333
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     317..333
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     352..372
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        354..372
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(399..461)
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
     misc_RNA        complement(469..547)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
ORIGIN
        1 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
       61 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      121 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      181 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      241 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      301 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
      361 cctcactaaa gggaacaaaa gctggagctc caccgcggtc tttctgtatc gcaaataagt
      421 gaaaattaaa aaagaaagat taaattggaa gttggatatg gaaaaaaagc accgactcgg
      481 tgccactttt tcaagttgat aacggactag ccttatttta acttgctatg ctgttttgaa
      541 tggttcctgc caatcgcagc tcccagagtt tcgttcccag ccgcgagcac cacagcgtac
      601 catgtgcgcc acgaggcctc aaacgtaaaa caatcgaaac gaaaagaaac agactatagg
      661 ggagtataga gacagccggc caataa
//
