LOCUS       sequence-109861-         836 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Constitutive expression of cas9 and inducible expression of lambda 
            RED and sgR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 836)
  AUTHORS   Jiang Y, Chen B, Duan C, Sun B, Yang J, Yang S
  TITLE     Multigene editing in the Escherichia coli genome using the 
            CRISPR-Cas9 system.
  JOURNAL   Appl Environ Microbiol. 2015 Jan 30. pii: AEM.04023-14.
  PUBMED    25636838
REFERENCE   2  (bases 1 to 836)
  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: "Appl 
            Environ Microbiol. 2015 Jan 30. pii: AEM.04023-14."
COMMENT     Sequence Label: pCas
FEATURES             Location/Qualifiers
     source          1..836
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    372..388
                     /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)."
     misc_RNA        418..493
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      complement(537..781)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
ORIGIN
        1 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
       61 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      121 taagttagcg cgaattgatc tggtttgaca gcttatcatc gactgcacgg tgcaccaatg
      181 cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg
      241 cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc
      301 ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc cggctcgtat
      361 gttgtgtgga attgtgagcg gataacaatt tcacacatct gctaatcctg ttaccaggtt
      421 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
      481 accgagtcgg tgcttttttt gagatctgtc catacccatg gattcttcgt ctgtttctac
      541 tggtattggc acaaacctga ttccaatttg agcaaggcta tgtgccatct cgatactcgt
      601 tcttaactca acagaagatg ctttgtgcat acagcccctc gtttattatt tatctcctca
      661 gccagccgct gtgctttcag tggatttcgg ataacagaaa ggccgggaaa tacccagcct
      721 cgctttgtaa cggagtagac gaaagtgatt gcgcctaccc ggatattatc gtgaggatgc
      781 gtcatcgcca ttgctcccca aatacaaaac caatttcagc cagtgcctcg tccatt
//
