LOCUS       sequence-103054-         901 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Streptomyces expression of codon-optimized Cas9 and custom gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 901)
  AUTHORS   Cobb RE, Wang Y, Zhao H
  TITLE     High-Efficiency Multiplex Genome Editing of Streptomyces Species 
            Using an Engineered CRISPR/Cas System.
  JOURNAL   ACS Synth Biol. 2014 Dec 8.
  PUBMED    25458909
REFERENCE   2  (bases 1 to 901)
  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: "ACS Synth 
            Biol. 2014 Dec 8."
COMMENT     Sequence Label: pCRISPomyces-2
FEATURES             Location/Qualifiers
     source          1..901
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(100..134)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     misc_RNA        complement(156..231)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    274..295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(310..315,316..333,334..340)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    348..364
                     /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)."
     primer_bind     353..375
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     372..388
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     372..388
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             384..680
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
     primer_bind     407..427
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        409..427
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(474..493)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(475..493)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(503..520)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(503..519)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(512..534)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
       61 cgccagcaac gcggcctttt tacggttcct ggcctctaga taaaaaacgc ccggcggcaa
      121 ccgagcgttc tgaacaaatc cagatggagt aaaaagcacc gactcggtgc cactttttca
      181 agttgataac ggactagcct tattttaact tgctatttct agctctaaaa catgtcttca
      241 ccggtggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
      301 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
      361 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
      421 ctaaagggaa caaaagctgg aggacctcga gggggggccc ggtacccaat tcgccctata
      481 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
      541 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      601 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
      661 acgcgccctg tagcggctga gaagacttgc gtatcccctt tcagatactc gcactaagaa
      721 ttgcaggaac gccccgatca tagcggtagc cgcccagatg ctgcaagcct tatctggcag
      781 ccgtataaaa aaagcaaccg aacaggccat tcacagaagt ttcacaccgc tcgccgaggg
      841 gctccgcacc cggtgcgcgc cgtacagcag cgcgcttctc ccagcatcgg ccagttcgcc
      901 c
//
