LOCUS       sequence-355194-        1851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a gRNA that targets next to PAM library, used for A. 
            chroococcum type I-E #2 system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1851)
  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 1851)
  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: pAc2_gRNA1
FEATURES             Location/Qualifiers
     source          1..1851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     147..166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        368..402
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     terminator      complement(563..597)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(668..1327)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1242..1261
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(1509..1851,1..246)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
       61 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      121 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      181 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      241 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      301 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtcgaga gcatcgatcc
      361 tagcatgttg acagctagct cagtcctagg tataatgcta gcgtcttccc cacgcccgtg
      421 ggggtgtttc aattctggcg aatcctttaa ttaactgacc agcgtcttcc ccacgcccgt
      481 gggggtgttt caaaaaaaaa ccccgcccct gacagggcgg ggtttttttt aaagaatcga
      541 tcgatcccga atgagtagac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
      601 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga
      661 tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
      721 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
      781 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
      841 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gaaacgaaaa
      901 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
      961 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     1021 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     1081 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     1141 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     1201 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     1261 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     1321 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     1381 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc aatcatgacc
     1441 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1501 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1561 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1621 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1681 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1741 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1801 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc c
//
