LOCUS       sequence-358542-        2255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LbCas12a gRNA containing BbsI restriction recognition sites in 
            spacer sequence for Golden Gate Assembly.
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: LbCas12a-gRNA_BbsI_CAM
FEATURES             Location/Qualifiers
     source          1..2255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(399..433)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(504..1163)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1078..1097
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(1402..1419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1536..2081)
                     /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(1616..1635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcttttctct tctcaaccct gaattcgcat ctagactgaa ctggccgata attgcagacg
       61 aacgcacagc taacaccacg tcgtccctat ctgctgccct aggtctatga gtggttgctg
      121 gataacttta cgggcatgca taaggctcgt ataatatatt cagggtttca aagattaaat
      181 aatttctact aagtgtagat tcgtcttcct tgtacttggt gaagacaact cggtaccaaa
      241 ttccagaaaa gaggcctccc gaaagggggg ccttttttcg ttttggtccg ctgagcagtt
      301 acagagatgt tacgaaccac tagtgcactg cagtacacct ttggtcgaaa aaaaaagccc
      361 gcactgtcag gtgcgggctt ttttctgtgt ttccaccaat aaaaaacgcc cggcggcaac
      421 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
      481 gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt
      541 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
      601 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
      661 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
      721 ttggctgaaa cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
      781 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
      841 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
      901 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
      961 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     1021 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     1081 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     1141 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     1201 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     1261 ccgatcaagc gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca
     1321 gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg
     1381 atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     1441 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
     1501 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
     1561 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
     1621 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
     1681 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
     1741 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
     1801 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
     1861 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     1921 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     1981 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
     2041 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc
     2101 agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca
     2161 gccccatacg atataagttg tttcagccaa aaaacttaag accgccggtc ttgtccacta
     2221 ccttgcagta atgcggtgga caggatcggc ggttt
//
