LOCUS       sequence-158791-         827 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA targeting YFP 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 827)
  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 827)
  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: pCas34
FEATURES             Location/Qualifiers
     source          1..827
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(13..47)
                     /label=BBa_J23113
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     misc_RNA        143..218
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             239..268
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             284..301
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(357..374)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      527..573
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(683..702)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(767..786)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ctttctctag tagctagcat aatccctagg actgagctag ccatcaggtt aacaaaaaag
       61 caggaatatc cctatcagtg atagacagac atccctatca gtgatagagt ataaagttaa
      121 ctggcatcgc cctcgccctc gcgttttaga gctagaaata gcaagttaaa ataaggctag
      181 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttgaagc ttgggcccga
      241 acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc atcatcatca
      301 ttgagtttaa acggtctcca gcttggctgt tttggcggat gagagaagat tttcagcctg
      361 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
      421 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
      481 ggtagtgtgg ggtctcccca tgcgagagta tggaactgcc aggcatcaaa taaaacggaa
      541 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga actacggggt
      601 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaac aataaaactg
      661 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct
      721 tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
      781 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgg
//
