LOCUS       sequence-103012-        1041 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Streptomyces expression of codon-optimized Cas9, tracrRNA, and 
            custom crRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1041)
  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 1041)
  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-1
FEATURES             Location/Qualifiers
     source          1..1041
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(105..152)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     protein_bind    225..246
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(261..266,267..284,285..291)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    299..315
                     /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     304..326
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     323..339
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     323..339
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             335..631
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
     primer_bind     358..378
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        360..378
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(425..444)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(426..444)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(454..471)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(454..470)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(463..485)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        complement(675..959)
                     /label=gapdh promoter
                     /note="glyceraldehyde-3-phosphate dehydrogenase promoter 
                     from Eggerthella lenta"
     terminator      complement(960..993)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
       61 aaaacgccag caacgcggcc tttttacggt tcctggcctc tagacaaaaa acccctcaag
      121 acccgtttag aggccccaag gggttatgct agttttggga ccattcaaaa cagcatagct
      181 ctatgtcttc accggtggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      241 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      301 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
      361 attaaccctc actaaaggga acaaaagctg gaggacctcg agggggggcc cggtacccaa
      421 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
      481 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      541 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      601 tggcgaatgg gacgcgccct gtagcggctg agaagacttt tgggaccatt caaaacagca
      661 tagctctaaa acccgcgtat cccctttcag atactcgcac taagaattgc aggaacgccc
      721 cgatcatagc ggtagccgcc cagatgctgc aagccttatc tggcagccgt ataaaaaaag
      781 caaccgaaca ggccattcac agaagtttca caccgctcgc cgaggggctc cgcacccggt
      841 gcgcgccgta cagcagcgcg cttctcccag catcggccag ttcgcccgat ccgtccgtgt
      901 cgcacagccg acggctgcgg taaggtgccc gtagacgcac gtccgaccga aggagcagct
      961 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagta aaaaaagcac
     1021 cgactcggtg ccactttttc a
//
