LOCUS       sequence-153687-         888 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  The reporter plasmid p11-LacY-wtx1 encodes the toxin CcdB gene under
            the inducible BAD promoter and a single copy of the endonuclease 
            cleavage site which is readily extendable to multiple copies..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 888)
  AUTHORS   Chen Z, Zhao H
  TITLE     A highly sensitive selection method for directed evolution of homing
            endonucleases.
  JOURNAL   Nucleic Acids Res. 2005 Oct 6;33(18):e154.
  PUBMED    16214805
REFERENCE   2  (bases 1 to 888)
  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: "Nucleic 
            Acids Res."; date: "2005-10-6"; volume: "33(18)"; pages: "e154"
COMMENT     Sequence Label: p11-LacY-wtx1
FEATURES             Location/Qualifiers
     source          1..888
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     72..91
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             181..486
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     365..384
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(581..598)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      751..837
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt
       61 cacactttgc tatgccatag catttttatc cataagatta gcggatccta cctgacgctt
      121 tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagcg attgaaaacg
      181 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
      241 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
      301 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
      361 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
      421 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttttgggga
      481 atataatcta gcattacgct agggataaca gggtaatatc acgctctaga catacggcat
      541 gcaagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag
      601 aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac
      661 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
      721 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
      781 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
      841 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgg
//
