LOCUS       sequence-160113-         898 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA targeting YFP +787 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 898)
  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 898)
  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: pCas59
FEATURES             Location/Qualifiers
     source          1..898
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             60..71
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(80..114)
                     /label=BBa_J23113
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     misc_RNA        210..285
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             306..335
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             351..368
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(424..441)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      594..640
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(750..769)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(834..853)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ttaagcagct ctaatgcgct gttaatcact ttacttttat ctaatctgga catctagtat
       61 ttctcctctt tctctagtag ctagcataat ccctaggact gagctagcca tcaggttaac
      121 aaaaaagcag gaatatccct atcagtgata gacagacatc cctatcagtg atagagtata
      181 aagttaactc ggcggtcacg aactccagcg ttttagagct agaaatagca agttaaaata
      241 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgaagcttg
      301 ggcccgaaca aaaactcatc tcagaagagg atctgaatag cgccgtcgac catcatcatc
      361 atcatcattg agtttaaacg gtctccagct tggctgtttt ggcggatgag agaagatttt
      421 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
      481 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
      541 cgccgatggt agtgtggggt ctccccatgc gagagtatgg aactgccagg catcaaataa
      601 aacggaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaact
      661 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgaacaat
      721 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
      781 aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa
      841 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaag
//
