LOCUS       sequence-163535-        1030 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  gRNA cassette-carrying vector with natMX marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1030)
  AUTHORS   Stovicek V, Borodina I, Forster J
  TITLE     CRISPR-Cas system enables fast and simple genome editing of 
            industrial Saccharomyces cerevisiae strains.
  JOURNAL   Metab Eng Commun. 2015 Mar 20;2:13-22. doi: 
            10.1016/j.meteno.2015.03.001. eCollection 2015 Dec.
  PUBMED    34150504
REFERENCE   2  (bases 1 to 1030)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab Eng 
            Commun."; date: "2015-03-20"; volume: "2:13-22. doi"; pages: " 
            10.1016/j.meteno.2015.03.001. eCollection 2015 Dec"
COMMENT     Sequence Label: pCfB2311 (SNR52p-gRNA.ADE2-SUP4t_natMX)
FEATURES             Location/Qualifiers
     source          1..1030
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(21..268)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     terminator      complement(274..293)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(298..372)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(393..661)
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     primer_bind     complement(679..699)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(679..697)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(718..734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(718..734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(731..753)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    742..758
                     /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)."
     promoter        complement(join(766..772,773..790,791..796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    811..832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(949..966)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gggcgaattg ggtaccggcc gcaaattaaa gccttcgagc gtcccaaaac cttctcaagc
       61 aaggttttca gtataatgtt acatgcgtac acgcgtctgt acagaaaaaa aagaaaaatt
      121 tgaaatataa ataacgttct taatactaac ataactataa aaaaataaat agggacctag
      181 acttcaggtt gtctaactcc ttccttttcg gttagagcgg atgtgggggg agggcgtgaa
      241 cgtaagcgtg acataactaa ttacatgact cgaagacata aaaaacaaaa aaagcaccac
      301 cgactcggtg ccactttttc aagttgataa cggactagcc ttattttaac ttgctatttc
      361 tagctctaaa actgtggata gtctctacaa ttgatcattt atctttcact gcggagaagt
      421 ttcgaacgcc gaaacatgcg caccaacttt cacttctaca gcgtttgacc aaaatctttt
      481 gaacagaaca ttgtagggtg tgaaaaaatg cgcaccttta ccgctagccc aagagggcac
      541 tacaaaatct agagttgtac ttcaaacgta catgtaatca ccttgtatat actcgaaaga
      601 aaacatcaag tttctgtata aatatgagtg aaagcataat catacattat cttttcaaag
      661 aagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
      721 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
      781 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
      841 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
      901 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
      961 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1021 acggttatcc
//
