LOCUS       sequence-142924-         980 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 980)
  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 980)
  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..980
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      16..87
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      103..130
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(297..885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(377..396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggatctcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
       61 atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg
      121 cctttctgcg tttataccta gggagacgtc ggcgttcggc tgcggcgagc ggtatcagct
      181 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      241 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      301 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      361 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      421 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      481 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      541 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      601 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      661 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      721 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      781 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      841 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      901 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      961 actagtgctt ggattctcac
//
