LOCUS       sequence-98862-e         721 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid encoding iCas9, tracrRNA and crRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 721)
  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 721)
  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: pCRCT
FEATURES             Location/Qualifiers
     source          1..721
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        56..324
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     CDS             complement(367..663)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
     primer_bind     513..535
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     527..544
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     528..544
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     554..573
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        554..572
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(620..640)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(620..638)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(659..675)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(659..675)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(672..694)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    683..699
                     /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)."
ORIGIN
        1 aactcacaaa ttagagcttc aatttaatta tatcagttat taccctgtgc tcgagtcttt
       61 gaaaagataa tgtatgatta tgctttcact catatttata cagaaacttg atgttttctt
      121 tcgagtatat acaaggtgat tacatgtacg tttgaagtac aactctagat tttgtagtgc
      181 cctcttgggc tagcggtaaa ggtgcgcatt ttttcacacc ctacaatgtt ctgttcaaaa
      241 gattttggtc aaacgctgta gaagtgaaag ttggtgcgca tgtttcggcg ttcgaaactt
      301 ctccgcagtg aaagataaat gatcgggttt tagagctatg ctgttttgaa tggtcccaaa
      361 gagacctcag ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
      421 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      481 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      541 cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga
      601 ggtcctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
      661 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
      721 g
//
