LOCUS       sequence-319745-        4372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Ec67 retron defense system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4372)
  AUTHORS   Gao L, Altae-Tran H, Bohning F, Makarova KS, Segel M, Schmid-Burgk 
            JL, Koob J, Wolf YI, Koonin EV, Zhang F
  TITLE     Diverse enzymatic activities mediate antiviral immunity in 
            prokaryotes.
  JOURNAL   Science. 2020 Aug 28;369(6507):1077-1084. doi: 
            10.1126/science.aba0372.
  PUBMED    32855333
REFERENCE   2  (bases 1 to 4372)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2020-08-28- 28"; volume: "369"; 
            issue: "6507"; pages: "1077-1084"
COMMENT     Sequence Label: pLG006 (Ec67 retron)
FEATURES             Location/Qualifiers
     source          1..4372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(311..328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(445..990)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(525..544)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3262..3309
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3529..4188)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4103..4122
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4189..4291)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg
       61 atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc
      121 ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg
      181 ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt
      241 catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat
      301 attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa
      361 tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag
      421 tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga
      481 aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      541 tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat
      601 tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag
      661 ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat
      721 ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag
      781 ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact
      841 gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg
      901 gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa
      961 gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat
     1021 ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
     1081 tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt
     1141 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     1201 taattgcgtt gcgcacgcgc tatcacgtaa aataggcaaa atacttctgg aaaacagaaa
     1261 gttgaagtga tatgttcata aacacgcatg taggcagatt tgttggttgt gaatcgcaac
     1321 cagtggcctt aatggcagga ggaatcgcct ccctaaaatc cttgattcag agctatacgg
     1381 caggtgtgct gtgcgaagga gtgcctgcat gcgtttctcc ttggcctttt ttcctctggg
     1441 atgaagaaga aatgacaaaa acatctaaac ttgacgcact tagggctgct acttcacgtg
     1501 aagacttggc taaaatttta gatgttaagt tggtattttt aactaacgtt ctatatagaa
     1561 tcggctcgga taatcaatac actcaattta caataccgaa gaaaggaaaa ggggtaagga
     1621 ctatttctgc acctacagac cggttgaagg acatccaacg aagaatatgt gacttacttt
     1681 ctgattgtag agatgagatc tttgctataa ggaaaattag taacaactat tcctttggtt
     1741 ttgagagggg aaaatcaata atcctaaatg cttataagca tagaggcaaa caaataatat
     1801 taaatataga tcttaaggat ttttttgaaa gctttaattt cggacgagtt agaggatatt
     1861 ttctttccaa tcaggatttt ttattaaatc ctgtggtggc aacgacactt gcaaaagctg
     1921 catgctataa tggaaccctc ccccagggaa gtccatgttc tcctattatc tcaaatctaa
     1981 tttgcaatat tatggatatg agattagcta aactggctaa aaaatatgga tgtacttata
     2041 gcagatatgc tgatgatata acaatttcta caaataaaaa tacatttccg ttagaaatgg
     2101 ctactgtgca acctgaaggg gttgttttgg gaaaagtttt ggtaaaagaa atagaaaact
     2161 ctggattcga aataaatgat tcaaagacta ggcttacgta taagacatca aggcaagaag
     2221 taacgggact tacagttaac agaatcgtta atattgatag atgttattat aaaaaaactc
     2281 gggcgttggc acatgctttg tatcgtacag gtgaatataa agtgccagat gaaaatggtg
     2341 ttttagtttc aggaggtctg gataaacttg aggggatgtt tggttttatt gatcaagttg
     2401 ataagtttaa caatataaag aaaaaactga acaagcaacc tgatagatat gtattgacta
     2461 atgcgacttt gcatggtttt aaattaaagt tgaatgcgcg agaaaaagca tatagtaaat
     2521 ttatttacta taaatttttt catggcaaca cctgtcctac gataattaca gaagggaaga
     2581 ctgatcggat atatttgaag gctgctttgc attctttgga gacatcatat cctgagttgt
     2641 ttagagaaaa aacagatagt aaaaagaaag aaataaatct taatatattt aaatctaatg
     2701 aaaagaccaa atatttttta gatctttctg ggggaactgc agatctgaaa aaatttgtag
     2761 agcgttataa aaataattat gcttcttatt atggttctgt tccaaaacag ccagtgatta
     2821 tggttcttga taatgataca ggtccaagcg atttacttaa ttttctgcgc aataaagtta
     2881 aaagctgccc agacgatgta actgaaatga gaaagatgaa atatattcat gttttctata
     2941 atttatatat agttctcaca ccattgagtc cttccggcga acaaacttca atggaggatc
     3001 ttttccctaa agatatttta gatatcaaga ttgatggtaa gaaattcaac aaaaataatg
     3061 atggagactc aaaaacggaa tatgggaagc atattttttc catgagggtt gttagagata
     3121 aaaagcggaa aatagatttt aaggcatttt gttgtatttt tgatgctata aaagatataa
     3181 aggaacatta taaattaatg ttaaatagct aatgaacagc cctaacgtta tgaacgctaa
     3241 ggctgatttt tcgcgcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     3301 ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc
     3361 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
     3421 acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc
     3481 aggcgtagca ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc
     3541 ctgccactca tcgcaatact gttgtaattc atttaacatt ctgccgacat ggaagccatc
     3601 acagacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     3661 atatttgccc atagtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     3721 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     3781 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     3841 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     3901 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     3961 cgccatacgg aactctggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     4021 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     4081 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     4141 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     4201 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     4261 gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca
     4321 gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt ga
//
