LOCUS       sequence-358154-        2323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing Cas12c sgRNA targeting GFP (1) for in vivo 
            interference assay in bacteria..
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: pCJH009_proD_Cas12c-sgRNA-GFP_CAM
FEATURES             Location/Qualifiers
     source          1..2323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(157..191)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(262..921)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     836..855
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(1160..1177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1294..1839)
                     /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(1374..1393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccagaaaaga ggcctcccga aaggggggcc ttttttcgtt ttggtccgct gagcagttac
       61 agagatgtta cgaaccacta gtgcactgca gtacaccttt ggtcgaaaaa aaaagcccgc
      121 actgtcaggt gcgggctttt ttctgtgttt ccaccaataa aaaacgcccg gcggcaaccg
      181 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
      241 ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
      301 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
      361 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
      421 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
      481 ggctgaaacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
      541 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
      601 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
      661 actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt
      721 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
      781 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
      841 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
      901 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
      961 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccc
     1021 gatcaagcgc tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt
     1081 gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat
     1141 acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
     1201 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
     1261 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
     1321 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     1381 ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac
     1441 cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg
     1501 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct
     1561 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac
     1621 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
     1681 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
     1741 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
     1801 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag
     1861 ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc
     1921 cccatacgat ataagttgtt tcagccaaaa aacttaagac cgccggtctt gtccactacc
     1981 ttgcagtaat gcggtggaca ggatcggcgg ttttcttttc tcttctcaac cctgaattcg
     2041 catctagact gaactggccg ataattgcag acgaacgcac agctaacacc acgtcgtccc
     2101 tatctgctgc cctaggtcta tgagtggttg ctggataact ttacgggcat gcataaggct
     2161 cgtataatat attcaggata ccacccgtgc atttctggat caatgatccg tacctcaatg
     2221 tccgggcgcg cagctagagc gacctgaaat ctgaaaagca ggattcaggt tgggtttgag
     2281 gtcccaattc ttgttgaaag caggattcac tcggtaccaa att
//
