LOCUS       sequence-319747-        6010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the ietAS defense system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6010)
  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 6010)
  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: pLG033 (ietAS)
FEATURES             Location/Qualifiers
     source          1..6010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      3365..3412
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3632..4291)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4206..4225
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4292..4394)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4786..4803)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4920..5465)
                     /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(5000..5019)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaatgataag agaaaggttg cttagagctc cccaggcgtt ggcaggagct caaagggctg
       61 gaggtgggat atctctgggc tcattaccgg tagatattga tagtcgactc aacactgttg
      121 atgtcagtta tcctaaatta gacagttcag agatttttct gcctgcagca atcagtaccc
      181 gtgttgaaga gtttatcact aatgttcaac gttatgatga gtttgttaaa gctgatgcag
      241 cattgccgag tcgtatgctc gtgtatggaa agccaggaac aggtaagact atgttatcta
      301 agtacatcgc tacccgctta gattttccac ttcttacagt gcgttgcgat actttgatta
      361 gtagtttatt gggacaaacc agcaaaaatc ttagacaggt tttcgattat gtaatgcaga
      421 ggccatcagt gcttttttta gacgaatttg atgctttagc tggagcaaga ggtaatgaga
      481 gagatatagg tgagcttcag cgagttgtca tttcactatt gcagaatatg gatgcggcat
      541 cagaggatac ggtaattatt gcctcaacta accatgagca acttctggat cctgcaatct
      601 ggaggcgatt tagcttcaga attccaatgc ctctgcctga catacatcag agagagttaa
      661 tttggaaaaa tcgtttaaag aatatgatat gtagcgatct agatttaagt gatttatcaa
      721 gaaaatcgga aggattatcc ggagcaataa ttgaacaggt gagcttggat gcacgtaggg
      781 atgcagttat tgaaggtgca agtgtgataa atcaccataa attgtatagg cgtttgtatc
      841 ttgctcaatc gcttatggaa ggtgtaaatt taagcactta cgaagatgaa attcgttggt
      901 tacgttctaa agataaaaaa ttattttcta tcagagttct tgctaatttg tacaaactta
      961 catcaagagt aatttcaaac attctgaagg agtcaggagc atatgagcag aaggggtaca
     1021 cagtttagta acgcaaaagt tacaaaccca atgttaagaa tccctttttc cagtagtgac
     1081 ttgggtgcaa tagtaaacgc tggcggtggg gcaaaggtat tggttgatgt aacagccgaa
     1141 tatagacaag ggctagtaag aaatttaaca accagtaaac attatttaga atccaaactt
     1201 tcagagtacc ctggaagctt gggtactttg gttttcaaat taagagacca gggaatagcc
     1261 aaaacgcata ggccgaacaa aattgctcaa gaggctggat tgcaaaatgc cggtcatgcc
     1321 aaaatagatg aaatgttggt tgctgctcat gccggctgtt ttgacgtatt agagtcagtc
     1381 attttacatc ggaatattaa agcgattttg gctaatctaa gcgcgattga gcgcattgaa
     1441 ccttgggatg agaataggaa ggttccagga ggcactgatg gtttgtttga atcatcaaac
     1501 atccttgtac gactatttga gtacacaggt gaagatgcaa cttacaacaa ctatgaaaac
     1561 gttatttcta tattagaaca acacggagtt aaatatgatg agattagaca aaaatgtggt
     1621 cttcccttat taaggataat ggatttatcc ccaaatgata gatatatatt agacattctc
     1681 attgattacc cgggtataag aacgttaatt cctgaaccaa aatattcagc attcccggtt
     1741 agtgtaagtg attctgttgg cattgaaaca aatagctttc ccgtaccatc agaagaatta
     1801 cccattgttg ctgtatttga cactggggta agccccatcg cggcaacaat tactccttgg
     1861 gtagtgagta gggaaacata cgtaattcct cctgatacga gttatgaaca tgggactatg
     1921 gtgtcttcat tgatatcagg cgctcatttt ttaaatgaca atcatccatg gattcctgat
     1981 acaaaatcta aaatccatga tgtttgtgcc ttagatgaaa atggatctta tatatcagat
     2041 ttaattctga ggctagcaga tgctgtaaat aaaagaccag atataaaagt ctggaatttg
     2101 tctttgggag gcggaccatg taatgagcag acgtttagtg attttgcgat ggagttagat
     2161 cggctcagcg ataaatttgg tattttgttt gtagttgctg caggtaatta tgtagatgaa
     2221 cctatacgta catggccaaa tcctgatccg cttggaggtg ctgatttaat ttcctctcct
     2281 ggagagtcag tccgagcact aacagttggt tcagtttctc atatggaagc taatgatgct
     2341 ttaagtgaaa ttggaacacc gacaccatat actcgtcgtg gccctgggcc tgtatttact
     2401 ccaaagccag atataatcca tgctggcggt ggggttcata gaccttggaa tgtaggagca
     2461 agcagtttaa aggtcgtagg gccagataat aggctttgct ctaattttgg tactagtttt
     2521 gctgctccaa ttgtggcaag tttagctgcg catacatggc agagaatagc cactaataca
     2581 gactttaatg tttcaccatc attgattaaa gcattattaa ttcattccgc tcaattatct
     2641 tctcctgatt actcgccaag tgaaagacgc tatttgggag cgggaattcc taatgaagtt
     2701 attgagacct tatatgatag tgatgatagg tttactctga ttttccaaac attcttggtt
     2761 cctggggtga ggtggagaaa ggataactat cccataccat cggcacttat tcaaaatgga
     2821 aaatttaaag gtgagattgt aattactgct gcatatgcac caccactgaa ccctaatgcc
     2881 ggcagtgaat atgttcgcgc gaacgtagag ctaagttttg gcttaattga gaataatact
     2941 ataaaaggaa aagtgcctat ggaaggagaa aacggtcaat ctggatatga gagagctcaa
     3001 attgagcatg gtggaaagtg gtcaccagta aaaattcatc gcaaggcatt taataaagga
     3061 attacttcgg gtaactgggc tcttcaggct aaaacaacgt tgagagcgaa tgaaccggcc
     3121 ttaatggagc ctttacctgt aactattgta gtaactttaa aatcattaga tggaaacaca
     3181 caagtttatg ctgatggcgt aagagcttta aatgctaata actgggctca ctatccattg
     3241 cctgctcgtg tgccagtttc cgtataacaa ctatataaat caaacccgct gtagcgggtt
     3301 tgatttattt gtgggtgtgt tttataaaaa taccgcccat acacaacaaa atacaacgca
     3361 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc
     3421 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     3481 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt
     3541 gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta gcaccaggcg
     3601 tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcaat
     3661 actgttgtaa ttcatttaac attctgccga catggaagcc atcacagacg gcatgatgaa
     3721 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatagtga
     3781 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
     3841 cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
     3901 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
     3961 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
     4021 aagggtgaac actatcccat atcaccagct caccgtcttt catcgccata cggaactctg
     4081 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
     4141 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
     4201 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
     4261 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
     4321 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     4381 ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa
     4441 cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgg
     4501 cgcaaagtgc gtcgggtgat gctgccaact tactgattta gtgtatgatg gtgtttttga
     4561 ggtgctccag tggcttctgt ttctatcagc tgtccctcct gttcagctac tgacggggtg
     4621 gtgcgtaacg gcaaaagcac cgccggacat cagcgctagc ggagtgtata ctggcttact
     4681 atgttggcac tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc
     4741 accggtgcgt cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc
     4801 gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt
     4861 cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt
     4921 ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
     4981 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
     5041 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
     5101 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
     5161 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5221 cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt
     5281 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
     5341 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
     5401 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
     5461 ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct
     5521 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt
     5581 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     5641 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgca atagaacgat
     5701 gaaggatgga agctacatat tctcggtact aagatttatt tttctgacac aaaatgacca
     5761 tttggcgtta cataatccca aaaaaacgta tcaaaaatct caaaatgcgt tacgattaga
     5821 gagtattttg attctgcgtg ctcatttttt gattgctgtg gctttttgtt gtgggagtgt
     5881 tgaatggatt atttatcaga agtgttaaaa atcattgaag gtgcaacaaa ggcaaatgct
     5941 tcgatggcta gtaattatgc tgggttgctg gcagataagc tcgaacaaaa aggggaggtc
     6001 aagcaagcca
//
