LOCUS       sequence-158816-         858 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA targeting YFP +52 from TSS; non-template strand; 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 858)
  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 858)
  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: pCas52
FEATURES             Location/Qualifiers
     source          1..858
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             57..68
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(77..111)
                     /label=BBa_J23113
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     CDS             complement(188..205)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_RNA        207..282
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             303..332
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             348..365
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(421..438)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      591..637
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(747..766)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(831..850)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 agcagctcta atgcgctgtt aatcacttta cttttatcta atctggacat ctagtatttc
       61 tcctctttct ctagtagcta gcataatccc taggactgag ctagccatca ggttaacaaa
      121 aaagcaggaa tatccctatc agtgatagac agacatccct atcagtgata gagtataaag
      181 ttaactgatg atgatgatga tgatgagttt tagagctaga aatagcaagt taaaataagg
      241 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttg aagcttgggc
      301 ccgaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat catcatcatc
      361 atcattgagt ttaaacggtc tccagcttgg ctgttttggc ggatgagaga agattttcag
      421 cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg
      481 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
      541 cgatggtagt gtggggtctc cccatgcgag agtatggaac tgccaggcat caaataaaac
      601 ggaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaactacg
      661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa
      721 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
      781 gtcttgctct aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
      841 ggctcgcgat aatgtcgg
//
