LOCUS       sequence-355195-        1852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a gRNA that targets next to PAM library, used for 
            Marinomonas sp. 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: pMs_gRNA1
FEATURES             Location/Qualifiers
     source          1..1852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..57
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        259..293
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     terminator      complement(455..489)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(560..1219)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1134..1153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(1401..1852,1..137)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
       61 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
      121 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
      181 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtcgag
      241 agcatcgatc ctagcatgtt gacagctagc tcagtcctag gtataatgct agcgcgttcc
      301 ccgagcacac ggggataaac cgaattctgg cgaatccttt aattaactga ccaggcgttc
      361 cccgagcaca cggggataaa ccgaaaaaaa aaccccgccc ctgacagggc ggggtttttt
      421 ttaaagaatc gatcgatccc gaatgagtag accaataaaa aacgcccggc ggcaaccgag
      481 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
      541 aagcgagctc gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
      601 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
      661 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
      721 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
      781 ctgaaacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
      841 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
      901 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
      961 tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca
     1021 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     1081 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     1141 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     1201 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     1261 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgcccga
     1321 tcaatcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1381 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1441 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1501 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1561 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1621 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1681 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1741 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1801 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg ca
//
