LOCUS       sequence-358543-        3557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing Cas12c tracrRNA and a GFP-targeting pre-crRNA 
            separately..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3557)
  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 3557)
  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: pCJH059_proD_Cas12c_DualRNA-GFP_CAM
FEATURES             Location/Qualifiers
     source          1..3557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(130..164)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(235..894)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     809..828
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(1133..1150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1267..1812)
                     /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(1347..1366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcgtctgc aattatcggc cagttcaggc catgagcggc ctcatttcta ctagtgcact
       61 gcagtacacc tttggtcgaa aaaaaaagcc cgcactgtca ggtgcgggct tttttctgtg
      121 tttccaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
      181 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc
      241 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
      301 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
      361 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
      421 taaatcaaaa ctggtgaaac tcacccaggg attggctgaa acgaaaaaca tattctcaat
      481 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
      541 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
      601 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
      661 tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
      721 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
      781 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
      841 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
      901 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
      961 attatggtga aagttggaac ctcttacgtg cccgatcaag cgctagcgga gtgtatactg
     1021 gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa
     1081 aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct tcctcgctca
     1141 ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg
     1201 gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa
     1261 gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca
     1321 gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct
     1381 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
     1441 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
     1501 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
     1561 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
     1621 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
     1681 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
     1741 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
     1801 aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa
     1861 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtttcagcca
     1921 aaaaacttaa gaccgccggt cttgtccact accttgcagt aatgcggtgg acaggatcgg
     1981 cggttttctt ttctcttctc aaccctgaat tcgcatctag actgaactgg ccgataattg
     2041 cagacgaacg cacagctaac accacgtcgt ccctatctgc tgccctaggt ctatgagtgg
     2101 ttgctggata actttacggg catgcataag gctcgtataa tatattcatc tagactcgag
     2161 atgctggcca aggaatttgg tttgtcggat gtaatgattg ggaagcgatg caaaaaactg
     2221 ggtgttgtaa aaccaaagcc gggattctgg gcgaaggtgg acgctgggaa agtgccacat
     2281 ccccaaggaa agaaagtagc cgggcagtaa ataacgcttt ggaatatgag acgtatctgt
     2341 agaatttttg gtatccagta aaaagagcaa aacgaaatga caaaacactc aatacccctt
     2401 cacgcattcc gcaactccgg cgcagacgcc agaaaatgga aaggccgcat tgctttgctg
     2461 gccaagcgcg gcaaagaaac gatgcgaaca ctgcagtttc cgctcgaaat gagcgagcct
     2521 gaagcactgc gccgtgcgcc caagaatcgc cgcaccaagg acaccaccgt aagcgagtat
     2581 cactgtgtct tttcagactg cggaaagaca atgcacgcgg atgagaatgc ggcagtgaat
     2641 atcggcggaa agttcattgc agatatcgaa aaatagtttg cacaaagtgc gagaaaagat
     2701 ataatacaca cataccaccc gtgcatttct ggatcaatga tccgtacctc aatgtccggg
     2761 cgcgcagcta gagcgacctg aaatctgcac gaaaaccggc gaaagccggt tttttgttgc
     2821 ctgtagtttg agtttatttc tttgcgcctg ggatggcaaa gccagctttc accgggcgac
     2881 cgccgatacg ctcaccttcg actgggatca caagccttgc cgtttccttg gcgagatagc
     2941 tgcgcagtgc caacacttcc gcatgcaaga catttttacg cctaccacct ttgtacacga
     3001 acttatagcg agctttggat ttttggatag gttcaatcga atttaggtag ggtatccaaa
     3061 gtgccgctcg accaccacgg atgtttttcc atgtccaaat agagtctgga tgttgaaatt
     3121 gtgagatttc catatgcaag ctgggtggct gttaaaaatg aatttcatga taatacataa
     3181 tgttatatta tgaacgactt cccccgcgtc actctcggta ccaaagacga acaataagac
     3241 gctgaaaagc gtcttttttc gttttggtcc ctgaaaggag gaactatatc cggatgagtc
     3301 aaccgttgta gctgtgttga catctggttt ggcagcggac caaaacgaaa aaaggccccc
     3361 ctttcgggag gcctcttttc tggaatttgg taccgaggga ttgaatcctg ctttcaacaa
     3421 gaattgggac ctcaaaccca acctgaatcc tgctcatatg aatatattat acgagcctta
     3481 tgcatgcccg taaagttatc cagcaaccac tcatagacct agggcagcag atagggacga
     3541 cgtggtgtta gctgtgc
//
