LOCUS       sequence-304756-        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"
     terminator      2152..2199
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2419..3078)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2993..3012
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3079..3181)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3573..3590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3707..4252)
                     /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(3787..3806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
       61 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgcacgcgc tatcacgtaa
      121 aataggcaaa atacttctgg aaaacagaaa gttgaagtga tatgttcata aacacgcatg
      181 taggcagatt tgttggttgt gaatcgcaac cagtggcctt aatggcagga ggaatcgcct
      241 ccctaaaatc cttgattcag agctatacgg caggtgtgct gtgcgaagga gtgcctgcat
      301 gcgtttctcc ttggcctttt ttcctctggg atgaagaaga aatgacaaaa acatctaaac
      361 ttgacgcact tagggctgct acttcacgtg aagacttggc taaaatttta gatgttaagt
      421 tggtattttt aactaacgtt ctatatagaa tcggctcgga taatcaatac actcaattta
      481 caataccgaa gaaaggaaaa ggggtaagga ctatttctgc acctacagac cggttgaagg
      541 acatccaacg aagaatatgt gacttacttt ctgattgtag agatgagatc tttgctataa
      601 ggaaaattag taacaactat tcctttggtt ttgagagggg aaaatcaata atcctaaatg
      661 cttataagca tagaggcaaa caaataatat taaatataga tcttaaggat ttttttgaaa
      721 gctttaattt cggacgagtt agaggatatt ttctttccaa tcaggatttt ttattaaatc
      781 ctgtggtggc aacgacactt gcaaaagctg catgctataa tggaaccctc ccccagggaa
      841 gtccatgttc tcctattatc tcaaatctaa tttgcaatat tatggatatg agattagcta
      901 aactggctaa aaaatatgga tgtacttata gcagatatgc tgatgatata acaatttcta
      961 caaataaaaa tacatttccg ttagaaatgg ctactgtgca acctgaaggg gttgttttgg
     1021 gaaaagtttt ggtaaaagaa atagaaaact ctggattcga aataaatgat tcaaagacta
     1081 ggcttacgta taagacatca aggcaagaag taacgggact tacagttaac agaatcgtta
     1141 atattgatag atgttattat aaaaaaactc gggcgttggc acatgctttg tatcgtacag
     1201 gtgaatataa agtgccagat gaaaatggtg ttttagtttc aggaggtctg gataaacttg
     1261 aggggatgtt tggttttatt gatcaagttg ataagtttaa caatataaag aaaaaactga
     1321 acaagcaacc tgatagatat gtattgacta atgcgacttt gcatggtttt aaattaaagt
     1381 tgaatgcgcg agaaaaagca tatagtaaat ttatttacta taaatttttt catggcaaca
     1441 cctgtcctac gataattaca gaagggaaga ctgatcggat atatttgaag gctgctttgc
     1501 attctttgga gacatcatat cctgagttgt ttagagaaaa aacagatagt aaaaagaaag
     1561 aaataaatct taatatattt aaatctaatg aaaagaccaa atatttttta gatctttctg
     1621 ggggaactgc agatctgaaa aaatttgtag agcgttataa aaataattat gcttcttatt
     1681 atggttctgt tccaaaacag ccagtgatta tggttcttga taatgataca ggtccaagcg
     1741 atttacttaa ttttctgcgc aataaagtta aaagctgccc agacgatgta actgaaatga
     1801 gaaagatgaa atatattcat gttttctata atttatatat agttctcaca ccattgagtc
     1861 cttccggcga acaaacttca atggaggatc ttttccctaa agatatttta gatatcaaga
     1921 ttgatggtaa gaaattcaac aaaaataatg atggagactc aaaaacggaa tatgggaagc
     1981 atattttttc catgagggtt gttagagata aaaagcggaa aatagatttt aaggcatttt
     2041 gttgtatttt tgatgctata aaagatataa aggaacatta taaattaatg ttaaatagct
     2101 aatgaacagc cctaacgtta tgaacgctaa ggctgatttt tcgcgcaata actagcataa
     2161 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag
     2221 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
     2281 gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc
     2341 tgccattcat ccgcttatta tcacttattc aggcgtagca ccaggcgttt aagggcacca
     2401 ataactgcct taaaaaaatt acgccccgcc ctgccactca tcgcaatact gttgtaattc
     2461 atttaacatt ctgccgacat ggaagccatc acagacggca tgatgaacct gaatcgccag
     2521 cggcatcagc accttgtcgc cttgcgtata atatttgccc atagtgaaaa cgggggcgaa
     2581 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
     2641 tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta
     2701 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
     2761 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
     2821 atcccatatc accagctcac cgtctttcat cgccatacgg aactctggat gagcattcat
     2881 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     2941 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     3001 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     3061 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     3121 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     3181 aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg
     3241 atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc
     3301 gggtgatgct gccaacttac tgatttagtg tatgatggtg tttttgaggt gctccagtgg
     3361 cttctgtttc tatcagctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca
     3421 aaagcaccgc cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga
     3481 tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag
     3541 cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc
     3601 gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc
     3661 aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc
     3721 gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag
     3781 gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg
     3841 cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg
     3901 acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt
     3961 cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat
     4021 gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc
     4081 atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca
     4141 gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc
     4201 ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca
     4261 tcttattaat cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc
     4321 acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat gc
//
