LOCUS       sequence-158792-         832 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA targeting cpcB expressed from aTc inducible system in pEZ03 in
            the NS1 locus with kanamycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 832)
  AUTHORS   Gordon GC, Korosh TC, Cameron JC, Markley AL, Begemann MB, Pfleger 
            BF
  TITLE     CRISPR interference as a titratable, trans-acting regulatory tool 
            for metabolic engineering in the cyanobacterium Synechococcus sp. 
            strain PCC 7002.
  JOURNAL   Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 
            10.1016/j.ymben.2016.07.007.
  PUBMED    27481676
REFERENCE   2  (bases 1 to 832)
  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: "Metab Eng. 
            2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 
            10.1016/j.ymben.2016.07.007."
COMMENT     Sequence Label: pCas35
FEATURES             Location/Qualifiers
     source          1..832
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             47..58
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(67..101)
                     /label=BBa_J23113
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     misc_RNA        197..272
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             293..322
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             338..355
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(411..428)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      complement(584..630)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(737..756)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 atgcgctgtt aatcacttta cttttatcta atctggacat ctagtatttc tcctctttct
       61 ctagtagcta gcataatccc taggactgag ctagccatca ggttaacaaa aaagcaggaa
      121 tatccctatc agtgatagac agacatccct atcagtgata gagtataaag ttaactgcta
      181 gaaatgaact cacctcgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
      241 atcaacttga aaaagtggca ccgagtcggt gctttttttg aagcttgggc ccgaacaaaa
      301 actcatctca gaagaggatc tgaatagcgc cgtcgatcat catcatcatc atcattgagt
      361 ttaaacggtc tccagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag
      421 attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg
      481 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
      541 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggccca
      601 gtctttcgac tgagcctttc gttttatctg ttgtttgtcg gtgaactacg gggtctgacg
      661 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa actgtctgct
      721 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctct
      781 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg gg
//
