LOCUS       sequence-75199-e         768 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses C. elegans optimized Cas9 from the eef-1A.1 (eft-3) 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 768)
  AUTHORS   Waaijers S, Portegijs V, Kerver J, Lemmens BB, Tijsterman M, van den
            Heuvel S, Boxem M
  TITLE     CRISPR/Cas9-Targeted Mutagenesis in Caenorhabditis elegans.
  JOURNAL   Genetics. 2013 Aug 26.
  PUBMED    23979586
REFERENCE   2  (bases 1 to 768)
  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: "Genetics. 
            2013 Aug 26."
COMMENT     Sequence Label: pMB63
FEATURES             Location/Qualifiers
     source          1..768
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(332..350)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(371..387)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(371..387)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(384..406)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    395..411
                     /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(419..425,426..443,444..449))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    464..485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(602..619)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tctctcgcta atttatttgc ttttttaatg ttattatttt atgacttttt atagtcactg
       61 aaaagtttgc atctgagtga agtgaatgct atcaaaatgt gattctgtct gatgtacttt
      121 cacaatctct cttcaattcc attttgaagt gctttaaacc cgaaaggttg agaaaaatgc
      181 gagcgctcaa atatttgtat tgtgttcgtt gagtgaccca acaaaaagag gaaactttat
      241 tgtgccgcca agaaaaaagt ctcattgcct aagagaaaca tatgtggttg gaaatcttcc
      301 agatcgtgga tcagagctcc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
      361 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      421 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      481 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      541 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      601 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      661 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      721 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctgg
//
