LOCUS       sequence-142925-        1037 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Nicking Cas9 derived from pWT Cas9 (Addgene #44250). Ampicillin 
            resistance marker, the tetracycline repressor (TetR), a ColE1 origin
            of replication and Streptococcus pyogenes Cas9 D10A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1037)
  AUTHORS   Standage-Beier K, Zhang Q, Wang X
  TITLE     Targeted Large-Scale Deletion of Bacterial Genomes Using 
            CRISPR-Nickases.
  JOURNAL   ACS Synth Biol. 2015 Nov 20;4(11):1217-25. doi: 
            10.1021/acssynbio.5b00132. Epub 2015 Oct 25.
  PUBMED    26451892
REFERENCE   2  (bases 1 to 1037)
  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"; doi: 
            "10.1021/acssynbio.5b00132"; journalName: "ACS Synth Biol"; date: 
            "2015-11-20- 20"; volume: "4"; issue: "11"; pages: "1217-25"
COMMENT     Sequence Label: pCas9(D10A)
FEATURES             Location/Qualifiers
     source          1..1037
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        69..124
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    105..123
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             142..153
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 agcagctcta atgcgctgtt aatcacttta cttttatcta atctagacat cattaattcc
       61 taatttttgt tgacactcta tcgttgatag agttatttta ccactcccta tcagtgatag
      121 agaaaagaat tcaaaagatc taaagaggag aaaggatcta tggataagaa atactcaata
      181 ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga tcactgatga atataaggtt
      241 ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt
      301 ataggggctc ttttatttga cagtggagag acagcggaag cgactcgtct caaacggaca
      361 gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt atctacagga gattttttca
      421 aatgagatgg cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc ttttttggtg
      481 gaagaagaca agaagcatga acgtcatcct atttttggaa atatagtaga tgaagttgct
      541 tatcatgaga aatatccaac tatctatcat ctgcgaaaaa aattggtaga ttctactgat
      601 aaagcggatt tgcgcttaat ctatttggcc ttagcgcata tgattaagtt tcgtggtcat
      661 tttttgattg agggagattt aaatcctgat aatagtgatg tggacaaact atttatccag
      721 ttggtacaaa cctacaatca attatttgaa gaaaacccta ttaacgcaag tggagtagat
      781 gctaaagcga ttctttctgc acgattgagt aaatcaagac gattagaaaa tctcattgct
      841 cagctccccg gtgagaagaa aaatggctta tttgggaatc tcattgcttt gtcattgggt
      901 ttgaccccta attttaaatc aaattttgat ttggcagaag atgctaaatt acagctttca
      961 aaagatactt acgatgatga tttagataat ttattggcgc aaattggaga tcaatatgct
     1021 gatttgtttt tggcagc
//
