LOCUS       sequence-358157-        2444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing a Cas12c sgRNA targeting GFP and another sgRNA 
            targeting RFP in one transcript for testing multiplexed repression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2444)
  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 2444)
  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: pCJH026_proD_Cas12c-multiplexing_CAM
FEATURES             Location/Qualifiers
     source          1..2444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(360..394)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(465..1124)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1039..1058
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(1363..1380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1497..2042)
                     /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(1577..1596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acctgaaatc tgaaaagcag gattcaggtt gggtttgagg tcccaattct tgttgaaata
       61 ccacccgtgc atttctggat caatgatccg tacctcaatg tccgggcgcg cagctagagc
      121 gacctgaaat ctgaaaagca ggattcaggt tgggtttgag ggcgagtagc gaagacgtag
      181 caggattcac tcggtaccaa attccagaaa agaggcctcc cgaaaggggg gccttttttc
      241 gttttggtcc gctgagcagt tacagagatg ttacgaacca ctagtgcact gcagtacacc
      301 tttggtcgaa aaaaaaagcc cgcactgtca ggtgcgggct tttttctgtg tttccaccaa
      361 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
      421 ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc
      481 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
      541 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
      601 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
      661 ctggtgaaac tcacccaggg attggctgaa acgaaaaaca tattctcaat aaacccttta
      721 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
      781 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
      841 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
      901 atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
      961 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     1021 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     1081 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     1141 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     1201 aagttggaac ctcttacgtg cccgatcaag cgctagcgga gtgtatactg gcttactatg
     1261 ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc
     1321 ggtgcgtcag cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct
     1381 acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct
     1441 ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc
     1501 cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga
     1561 aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct
     1621 cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat
     1681 tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga
     1741 accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1801 ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag
     1861 tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc
     1921 ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc
     1981 cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc
     2041 aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa tttatctctt
     2101 caaatgtagc acctgaagtc agccccatac gatataagtt gtttcagcca aaaaacttaa
     2161 gaccgccggt cttgtccact accttgcagt aatgcggtgg acaggatcgg cggttttctt
     2221 ttctcttctc aaccctgaat tcgcatctag actgaactgg ccgataattg cagacgaacg
     2281 cacagctaac accacgtcgt ccctatctgc tgccctaggt ctatgagtgg ttgctggata
     2341 actttacggg catgcataag gctcgtataa tatattcagg ataccacccg tgcatttctg
     2401 gatcaatgat ccgtacctca atgtccgggc gcgcagctag agcg
//
