LOCUS       sequence-355157-        1852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a gRNA that targets next to PAM library, used for L. 
            mobilis type I-E system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1852)
  AUTHORS   Wimmer F, Mougiakos I, Englert F, Beisel CL
  TITLE     Rapid cell-free characterization of multi-subunit CRISPR effectors 
            and transposons.
  JOURNAL   Mol Cell. 2022 Feb 16. pii: S1097-2765(22)00102-2. doi: 
            10.1016/j.molcel.2022.01.026.
  PUBMED    35216669
REFERENCE   2  (bases 1 to 1852)
  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"; journalName: "Mol Cell. 
            2022 Feb 16. pii: S1097-2765(22)00102-2. doi: 
            10.1016/j.molcel.2022.01.026."
COMMENT     Sequence Label: pLm_gRNA1
FEATURES             Location/Qualifiers
     source          1..1852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     141..160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        362..396
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     terminator      complement(558..592)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(663..1322)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1237..1256
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(1504..1852,1..240)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
       61 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      121 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      181 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      241 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      301 ttctttcctg cgttatcccc tgattctgtg gataaccgtc gagagcatcg atcctagcat
      361 gttgacagct agctcagtcc taggtataat gctagcgtgg tccccgcgct ggcggggatg
      421 aaccgaattc tggcgaatcc tttaattaac tgaccaggtg gtccccgcgc tggcggggat
      481 gaaccgaaaa aaaaaccccg cccctgacag ggcggggttt tttttaaaga atcgatcgat
      541 cccgaatgag tagaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca
      601 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgatatca
      661 aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
      721 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
      781 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
      841 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgaaac gaaaaacata
      901 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
      961 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     1021 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     1081 tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg
     1141 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     1201 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     1261 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     1321 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
     1381 cttatttcat tatggtgaaa gttggaacct cttacgtgcc cgatcaatca tgaccaaaat
     1441 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1501 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1561 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1621 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     1681 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1741 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1801 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tg
//
