LOCUS       sequence-358160-        3633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing Cas12c tracrRNA and GFP-targeting crRNA 
            separately, with the crRNA ending with a HDV ribozyme sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3633)
  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 3633)
  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: pCJH065_proD_Cas12c_DualRNA-GFP+HDVrz_CAM
FEATURES             Location/Qualifiers
     source          1..3633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(126..160)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(231..890)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     805..824
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(1129..1146)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1263..1808)
                     /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(1343..1362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtctgcaatt atcggccagt tcaggccatg agcggcctca tttctactag tgcactgcag
       61 tacacctttg gtcgaaaaaa aaagcccgca ctgtcaggtg cgggcttttt tctgtgtttc
      121 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
      181 aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg
      241 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
      301 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
      361 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
      421 tcaaaactgg tgaaactcac ccagggattg gctgaaacga aaaacatatt ctcaataaac
      481 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
      541 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
      601 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
      661 attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
      721 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
      781 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
      841 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
      901 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
      961 tggtgaaagt tggaacctct tacgtgcccg atcaagcgct agcggagtgt atactggctt
     1021 actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc
     1081 tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga
     1141 ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga
     1201 tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg
     1261 tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg
     1321 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg
     1381 cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg
     1441 tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat
     1501 gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1561 caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg
     1621 agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact
     1681 gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa
     1741 aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac
     1801 gatctcaaga agatcatctt attaatcaga taaaatattt ctagatttca gtgcaattta
     1861 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgttt cagccaaaaa
     1921 acttaagacc gccggtcttg tccactacct tgcagtaatg cggtggacag gatcggcggt
     1981 tttcttttct cttctcaacc ctgaattcgc atctagactg aactggccga taattgcaga
     2041 cgaacgcaca gctaacacca cgtcgtccct atctgctgcc ctaggtctat gagtggttgc
     2101 tggataactt tacgggcatg cataaggctc gtataatata ttcatctaga ctcgagatgc
     2161 tggccaagga atttggtttg tcggatgtaa tgattgggaa gcgatgcaaa aaactgggtg
     2221 ttgtaaaacc aaagccggga ttctgggcga aggtggacgc tgggaaagtg ccacatcccc
     2281 aaggaaagaa agtagccggg cagtaaataa cgctttggaa tatgagacgt atctgtagaa
     2341 tttttggtat ccagtaaaaa gagcaaaacg aaatgacaaa acactcaata ccccttcacg
     2401 cattccgcaa ctccggcgca gacgccagaa aatggaaagg ccgcattgct ttgctggcca
     2461 agcgcggcaa agaaacgatg cgaacactgc agtttccgct cgaaatgagc gagcctgaag
     2521 cactgcgccg tgcgcccaag aatcgccgca ccaaggacac caccgtaagc gagtatcact
     2581 gtgtcttttc agactgcgga aagacaatgc acgcggatga gaatgcggca gtgaatatcg
     2641 gcggaaagtt cattgcagat atcgaaaaat agtttgcaca aagtgcgaga aaagatataa
     2701 tacacacata ccacccgtgc atttctggat caatgatccg tacctcaatg tccgggcgcg
     2761 cagctagagc gacctgaaat ctgcacgaaa accggcgaaa gccggttttt tgttgcctgt
     2821 agtttgagtt tatttctttg cgcctgggat ggcaaagcca gctttcaccg ggcgaccgcc
     2881 gatacgctca ccttcgactg ggatcacaag ccttgccgtt tccttggcga gatagctgcg
     2941 cagtgccaac acttccgcat gcaagacatt tttacgccta ccacctttgt acacgaactt
     3001 atagcgagct ttggattttt ggataggttc aatcgaattt aggtagggta tccaaagtgc
     3061 cgctcgacca ccacggatgt ttttccatgt ccaaatagag tctggatgtt gaaattgtga
     3121 gatttccata tgcaagctgg gtggctgtta aaaatgaatt tcatgataat acataatgtt
     3181 atattatgaa cgacttcccc cgcgtcactc tcggtaccaa agacgaacaa taagacgctg
     3241 aaaagcgtct tttttcgttt tggtccctga aaggaggaac tatatccgga tgagtcaacc
     3301 gttgtagctg tgttgacatc tggtttggca gcggaccaaa acgaaaaaag gccccccttt
     3361 cgggaggcct cttttctgga atttggtacc gagggattga atcctgctgg ctctccctta
     3421 gccatccgag tttcctcgga tgcccaggtc ggaccgcgag gaggtggaga tgccatgccg
     3481 acccttcaac aagaattggg acctcaaacc caacctgaat cctgctcata tgaatatatt
     3541 atacgagcct tatgcatgcc cgtaaagtta tccagcaacc actcatagac ctagggcagc
     3601 agatagggac gacgtggtgt tagctgtgcg ttc
//
