LOCUS       sequence-358161-        2323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas12c sgRNA containing BbsI restriction recognition sites in spacer
            sequence for Golden Gate Assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2323)
  AUTHORS   Huang CJ, Adler BA, Doudna JA
  TITLE     A naturally DNase-free CRISPR-Cas12c enzyme silences gene 
            expression.
  JOURNAL   Mol Cell. 2022 Jun 2;82(11):2148-2160.e4. doi: 
            10.1016/j.molcel.2022.04.020.
  PUBMED    35659325
REFERENCE   2  (bases 1 to 2323)
  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"; doi: 
            "10.1016/j.molcel.2022.04"; journalName: "Mol Cell"; date: 
            "2022-06-2- 2"; volume: "82"; issue: "11"; pages: "2148-2160.e4"
COMMENT     Sequence Label: Cas12c-sgRNA_BbsI_CAM
FEATURES             Location/Qualifiers
     source          1..2323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(301..335)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(406..1065)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     980..999
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(1304..1321)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1438..1983)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1518..1537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accacccgtg catttctgga tcaatgatcc gtacctcaat gtccgggcgc gcagctagag
       61 cgacctgaaa tctgaaaagc aggattcagg ttgggtttga ggtcgtcttc cgaagactta
      121 gcaggattca ctcggtacca aattccagaa aagaggcctc ccgaaagggg ggcctttttt
      181 cgttttggtc cgctgagcag ttacagagat gttacgaacc actagtgcac tgcagtacac
      241 ctttggtcga aaaaaaaagc ccgcactgtc aggtgcgggc ttttttctgt gtttccacca
      301 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
      361 attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg
      421 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
      481 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
      541 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
      601 actggtgaaa ctcacccagg gattggctga aacgaaaaac atattctcaa taaacccttt
      661 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
      721 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
      781 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
      841 catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
      901 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
      961 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     1021 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     1081 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     1141 aaagttggaa cctcttacgt gcccgatcaa gcgctagcgg agtgtatact ggcttactat
     1201 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     1261 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     1321 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     1381 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     1441 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     1501 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     1561 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     1621 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     1681 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1741 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     1801 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     1861 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     1921 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     1981 caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca atttatctct
     2041 tcaaatgtag cacctgaagt cagccccata cgatataagt tgtttcagcc aaaaaactta
     2101 agaccgccgg tcttgtccac taccttgcag taatgcggtg gacaggatcg gcggttttct
     2161 tttctcttct caaccctgaa ttcgcatcta gactgaactg gccgataatt gcagacgaac
     2221 gcacagctaa caccacgtcg tccctatctg ctgccctagg tctatgagtg gttgctggat
     2281 aactttacgg gcatgcataa ggctcgtata atatattcag gat
//
