LOCUS       sequence-355198-        1852 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 #3 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: pAc3_gRNA1
FEATURES             Location/Qualifiers
     source          1..1852
                     /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(564..598)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(669..1328)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1243..1262
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(1510..1852,1..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 gcgtgttccc cgcgcctgcg
      421 gggatgaacc gaattctggc gaatccttta attaactgac caggtgttcc ccgcgcctgc
      481 ggggatgaac cgaaaaaaaa accccgcccc tgacagggcg gggttttttt taaagaatcg
      541 atcgatcccg aatgagtaga ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
      601 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
      661 atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
      721 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
      781 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
      841 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgaaacgaaa
      901 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
      961 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     1021 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     1081 accagctcac cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca
     1141 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     1201 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     1261 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     1321 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     1381 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcccgat caatcatgac
     1441 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1501 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1561 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1621 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1681 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1741 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1801 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc cc
//
