LOCUS       sequence-358159-        2255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing Cas12a guide RNA 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 2255)
  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 2255)
  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: pCJH031_proD_LbCas12a-gRNA-GFP_CAM
FEATURES             Location/Qualifiers
     source          1..2255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(238..272)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(343..1002)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     917..936
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(1241..1258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1375..1920)
                     /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(1455..1474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agggtttcaa agattaaata atttctacta agtgtagata ctggagttgt cccaattctt
       61 gttgaatctc ggtaccaaat tccagaaaag aggcctcccg aaaggggggc cttttttcgt
      121 tttggtccgc tgagcagtta cagagatgtt acgaaccact agtgcactgc agtacacctt
      181 tggtcgaaaa aaaaagcccg cactgtcagg tgcgggcttt tttctgtgtt tccaccaata
      241 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
      301 actggatcta tcaacaggag tccaagcgag ctcgatatca aattacgccc cgccctgcca
      361 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacaaac
      421 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
      481 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
      541 ggtgaaactc acccagggat tggctgaaac gaaaaacata ttctcaataa accctttagg
      601 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
      661 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
      721 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
      781 acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
      841 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
      901 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
      961 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
     1021 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     1081 gttggaacct cttacgtgcc cgatcaagcg ctagcggagt gtatactggc ttactatgtt
     1141 ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg
     1201 tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac
     1261 gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg
     1321 aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca
     1381 taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa
     1441 cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc
     1501 tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc
     1561 cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac
     1621 cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1681 aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc
     1741 ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct
     1801 ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc
     1861 tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa
     1921 gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt tatctcttca
     1981 aatgtagcac ctgaagtcag ccccatacga tataagttgt ttcagccaaa aaacttaaga
     2041 ccgccggtct tgtccactac cttgcagtaa tgcggtggac aggatcggcg gttttctttt
     2101 ctcttctcaa ccctgaattc gcatctagac tgaactggcc gataattgca gacgaacgca
     2161 cagctaacac cacgtcgtcc ctatctgctg ccctaggtct atgagtggtt gctggataac
     2221 tttacgggca tgcataaggc tcgtataata tattc
//
