LOCUS       sequence-355197-        1852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a gRNA that targets next to PAM library, used for E. 
            haloalkaliphila 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: pEh_gRNA1
FEATURES             Location/Qualifiers
     source          1..1852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        372..406
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     terminator      complement(568..602)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(673..1332)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1247..1266
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(1514..1852,1..250)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
       61 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      301 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtc gagagcatcg
      361 atcctagcat gttgacagct agctcagtcc taggtataat gctagcccgt tccccgcagg
      421 cgcggggatg aaccgaattc tggcgaatcc tttaattaac tgaccagccg ttccccgcag
      481 gcgcggggat gaaccgaaaa aaaaaccccg cccctgacag ggcggggttt tttttaaaga
      541 atcgatcgat cccgaatgag tagaccaata aaaaacgccc ggcggcaacc gagcgttctg
      601 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag
      661 ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
      721 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
      781 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
      841 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgaaac
      901 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
      961 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     1021 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     1081 tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg
     1141 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     1201 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
     1261 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     1321 ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     1381 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc cgatcaatca
     1441 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1501 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1561 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1621 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1681 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1741 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1801 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca ca
//
