LOCUS       sequence-141189-         926 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 926)
  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 926)
  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..926
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(16..36)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(16..34)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(55..71)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(55..71)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(68..90)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    79..95
                     /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(103..109,110..127,128..133))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    148..169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      242..261
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        263..662
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             674..697
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             701..721
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
       61 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      121 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
      181 ctcactgccc gctttccacc ggtggtctct agagctatgc tgttttgaat ggtcccaaaa
      241 ctttttttgt tttttatgtc tgcatagctt caaaatgttt ctactccttt tttactcttc
      301 cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac
      361 taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa
      421 aagaaaaaag aggccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta
      481 tcacgtttct ttttcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc
      541 attgatattt aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg
      601 ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta
      661 agttttctag atggattata aagatgacga tgacaaacct ccaaaaaaga agagaaaggt
      721 cgataagaaa tactcaatag gcttagatat cggcacaaat agcgtcggat gggcggtgat
      781 cactgatgaa tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca
      841 cagtatcaaa aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc
      901 gactcgtctc aaacggacag ctcgta
//
