LOCUS       sequence-342945-        1852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes E. coli type I-E single spacer array.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1852)
  AUTHORS   Wimmer F, Englert F, Beisel CL
  TITLE     A TXTL-Based Assay to Rapidly Identify PAMs for CRISPR-Cas Systems 
            with Multi-Protein Effector Complexes.
  JOURNAL   Methods Mol Biol. 2022;2433:391-411. doi: 
            10.1007/978-1-0716-1998-8_24.
  PUBMED    34985758
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: "Methods Mol
            Biol."; date: "2022"; volume: "2433:391-411. doi"; pages: " 
            10.1007/978-1-0716-1998-8_24"
COMMENT     Sequence Label: pEc-crRNA2
FEATURES             Location/Qualifiers
     source          1..1852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(153..741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(233..252)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             923..1582
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(989..1008)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1653..1687
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     promoter        complement(join(1849..1852,1..31))
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 gcattatacc taggactgag ctagctgtca acatgctagg atcgatgctc tcgacggtta
       61 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      121 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      181 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      241 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      301 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      361 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      421 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      481 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      541 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
      601 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      661 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
      721 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
      781 tggaacgaaa actcacgtta agggattttg gtcatgattg atcgggcacg taagaggttc
      841 caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt
      901 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
      961 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     1021 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     1081 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     1141 tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     1201 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     1261 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     1321 tttccctaaa gggtttattg agaatatgtt tttcgtttca gccaatccct gggtgagttt
     1381 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     1441 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     1501 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     1561 tgagtggcag ggcggggcgt aatttgatat cgagctcgct tggactcctg ttgatagatc
     1621 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt
     1681 tttttattgg tctactcatt cgggatcgat cgattcttta aaaaaaaccc cgccctgtca
     1741 ggggcggggt ttttttttcg gtttatcccc gctggcgcgg ggaactcaca cgcacggtgt
     1801 tagctcagca tgcccagcgc ggtttatccc cgctggcgcg gggaactcgc ta
//
