LOCUS       sequence-319749-        7298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the AVAST type 4 defense system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7298)
  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 7298)
  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: pLG020 (AVAST type 4)
FEATURES             Location/Qualifiers
     source          1..7298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..546)
                     /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(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5744..5791
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(6011..6670)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6585..6604
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6671..6773)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(7165..7182)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
      121 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
      181 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
      241 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      301 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
      361 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
      421 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
      481 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
      541 tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc
      601 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag
      661 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
      721 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc agcgcagctg
      781 acaaagattg accgtgagcg ctctgatgga gaaagacgat agttgctgag tacgatatcg
      841 agggtacatt tctctgtgta ggggtagtta tttacaaaaa aataggagaa taattaaatg
      901 gtcaaaccaa actgggataa ctttaaagct aaatttagtg agaatcctca aggtaatttt
      961 gagtggtttt gctacttgtt gttctgtcaa gaattcaaaa tgcccgcagg tatatttaga
     1021 tataagaatc aatctggtat cgaaactaat ccaataacca aagataatga aattatcggt
     1081 tggcaatcta aattctatga cacaaaattg tcggataaca aagctgatct tatagaaatg
     1141 attgagaaaa gcaaaaaggc ttatccagga ttaagtaaaa tcattttcta tactaatcaa
     1201 gagtgggggc aggggagaaa gtcccatgaa cctgaaggcg ataagaacgc tgataattat
     1261 ttggaaactg tcggaaatag taacgatccc aaaataaaaa ttgaagttga tcagaaagca
     1321 tatgagtcgg gtatcgaaat agtatggaga gttgctagtt tttttgaatc accgtttgta
     1381 atagttgaga atgaaaagat tgctaaacat ttcttctccc ttaatgaaag catctttgat
     1441 ttattagaag aaaagcgcaa gcacacagaa aatgttttat atgaaattca aaccaatata
     1501 gagttcaaag acagaagtat tgaaattgac agacgacatt gcatagaact tctacatgag
     1561 aatctagttc agaaaaaaat tgtcatcgtc agcggagaag gtggggttgg aaaaacagca
     1621 gttatcaaaa aaatttatga agcagaaaaa caatacactc ctttctatgt ctttaaggct
     1681 agcgagttta aaaaggacag cattaatgag ttattcggtg cgcatggctt agacgatttc
     1741 tctaatgctc atcaagacga attacgtaaa gtcatagtcg tagattctgc tgaaaagctt
     1801 ttagaactga ccaatatcga tccttttaaa gaattcctga ctgttttaat aaaggataaa
     1861 tggcaggttg ttttcacaac ccgtaacaat tacttggcag atctgaacta tgctttcata
     1921 gatatttata agataactcc tggaaactta gtaataaaga accttgaacg cggcgagcta
     1981 atagagttat ctgataacaa tggatttagc cttcctcaag atgttcgatt attagaacta
     2041 atcaaaaatc cattttatct aagtgaatat ttgaggttct ataccggtga aagcatcgat
     2101 tatgtgagct tcaaagaaaa gctatggaat aagattatcg tcaaaaataa accttctcgg
     2161 gagcagtgtt tcttagcgac tgcttttcag cgggctagtg agggccaatt ttttgtctcc
     2221 ccggcatgtg atactggaat tttagatgag ttagttaaag acggaattgt cggctatgaa
     2281 gctgctggtt acttcattac acatgatata tacgaggaat gggcattaga aaagaaaatt
     2341 tctgtcgatt atatccgtaa agcgaacaat aacgagttct tcgaaaaaat aggagaatca
     2401 cttcctgttc gccgtagttt tcggaattgg atatctgaac gattgctttt agatgaccag
     2461 tccataaagc cttttatcgc agaaatagtc tgtggagaag gaatatcaaa tttttggaaa
     2521 gacgagttat gggtagctgt ccttctttcc gacaattcaa gcatattttt taattacttt
     2581 aaaagatatt tacttagtag tgaccagaat ctattaaaaa gacttacttt cttattgagg
     2641 cttgcttgca aggacgttga ttacgatctg cttaaacagt taggtgtaag taattcagat
     2701 ctgctttcca ttaaatatgt tcttactaag cctaagggaa ctggttggca gagtgtgatc
     2761 caatttatct atgaaaattt agatgaaata gggatcagaa atattaattt tatacttcct
     2821 gtgattcagg agtggaatca aagaaacaaa gtgggtgaaa cgactcgatt atctagtttg
     2881 atagctctaa aatattatca atggactata gatgaggatg tctatttatc cggaagggat
     2941 aatgagaaaa atattctgca tacgattctt catggggcgg ccatgattaa acctgaaatg
     3001 gaagaggttt tagttaaggt tcttaaaaat aggtggaaag agcatggtac cccatatttc
     3061 gaccttatga ccttaatcct tactgactta gattcatatc cggtttgggc atctctcccg
     3121 gaatatgttc tacaattggc agatctgttc tggtatcggc cacttaaaga aacaggcgaa
     3181 cgttatcaca gtatggatat tgaagatgag ttcggtctat ttaggtctca tcacgactat
     3241 tatccagaaa gtccatatca gactcctata tattggttac tacaatcaca gttcaaaaaa
     3301 acaatagact ttattcttga ttttacgaac aagacaacga tatgttttgc ccactcccat
     3361 tttgctaaaa acgaaattga agaagtagat gtctttattg aagaaggaaa gtttataaag
     3421 caatatatat gcaatcgtct gtggtgctca taccgaggaa cacaggtctc tacctactta
     3481 ctttcatcaa ttcatatggc attggaaaag ttttttcttg agaattttaa aaatgcagac
     3541 tcgaaagtgt tggaaagttg gcttcttttc ttgttaagaa ataccaagtc agcttctatt
     3601 tctgcagtag ttacgagtat tgtacttgca ttccctgaga agacattcaa tgtagctaaa
     3661 gtactattcc aaacaaagga cttcttccgt tttgatatga atcgaatggt tctagacaga
     3721 acacataaaa gttcattaat ctccctcagg gatggctttg gcggtacaga ttacagaaac
     3781 tctttgcacg aagaagatag aattaaagct tgcgatgatg tgcatagaaa tacttatctt
     3841 gaaaatcttg ccttgcatta tcaaattttc aggagtgaaa atgtaacgga gaaagatgcc
     3901 attgaaaggc aacaagtgct ctgggatatt ttcgacaaat actataatca gcttccagat
     3961 gaagctcaag aaactgaagc cgataagacg tggaggctct gcttggcaag aatggatcgg
     4021 cgaaagatga aaataactac caaggagaaa gatgaaggga ttgagatatc attcaatcct
     4081 gagattgacc ctaaactaaa gcaatatagt gaggaagcaa taaagaaaaa ctccgagcat
     4141 atgaagtatg taacgctgaa actatgggca agctataaaa gagaaaagga tgaacgttat
     4201 aagaattatg gaatgtatga ggacaatccg caaattgctt tacaagagac caaagaaata
     4261 ataaaaaagc ttaatgagga agggggtgaa gatttcagac tattaaatgg taatatacca
     4321 gcagacgttt gttctgtatt actgttagat tattttaatc agttgaataa tgaagagaga
     4381 gaatactgta aagatattgt tctagcgtat tctaaacttc cgttgaagga aggctataat
     4441 tatcaggtac aagatggaac aacctcggca atttcagcct tacccgtgat ttatcataat
     4501 tatccaatgg aaagggagac tataaaaaca atattacttt tgacactgtt taatgaccac
     4561 tctattggaa tggcaggtgg gcgctactca gtatttccta gtatggtgat tcataaatta
     4621 tggctagact attttgatga tatgcagtcc ctattgtttg gttttttgat tttaaagcca
     4681 aaatatgtaa tcctttcaag aaaaatcatt catgaaagtt atcgtcaagt agactatgac
     4741 attaaaaaaa taaatattaa taaggtgttt ttaaataact ataagcattg catatcaaat
     4801 gtcatcgata ataaaatatc tatagatgat ttgggaagta tggataaagt tgatctacat
     4861 attttgaaca cagctttcca attaattcca gttgatactg ttaatattga acataagaaa
     4921 ttggtttcct taattgttaa aagattttct acaagcctat tgtcaagtgt tcgagaagat
     4981 agagttgatt acgctcttcg gcagtctttc ttggaaagat ttgcctactt tacgcttcat
     5041 gcgcccgtga gcgatattcc cgattatata aaaccttttc ttgatggttt caacggttca
     5101 gagcctattt cagagttatt taaaaaattt attctcgtcg aagatagatt aaatacttac
     5161 gccaaatttt ggaaggtttg ggatttgttt tttgataaag tggttacttt gtgcaaggat
     5221 ggagataggt attggtatgt agataaaatt ataaaaagtt acctttttgc tgaatctcca
     5281 tggaaagaaa actctaatgg ttggcacaca tttaaagata gcaatagtca attcttttgc
     5341 gatgtatcta ggactatggg ccattgccct tcaactttat attctcttgc caaatctttg
     5401 aataacattg ccagttgcta tcttaatcaa ggtataactt ggctttcaga aatattgtcg
     5461 gttaataaaa agctatggga aaagaaattg gaaaatgata ctgtttatta tttggaatgt
     5521 ttggttaggc ggtatattaa caatgagcgt gagcgaatta gacgaaccaa acagttgaaa
     5581 caagaggtct tagtaatatt ggattttttg gtagagaaag gatcggttgt tggttatatg
     5641 tcacgggaaa atattctgtg atgtagttga aaataataat tttaatgaga gcttttccaa
     5701 tttaggctcc agggattgga gcctttttat tatcgcgcaa taactagcat aaccccttgg
     5761 ggcctctaaa cgggtcttga ggggtttttt gctgaaacct caggcatttg agaagcacac
     5821 ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat
     5881 ttaacgaccc tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt tctgccattc
     5941 atccgcttat tatcacttat tcaggcgtag caccaggcgt ttaagggcac caataactgc
     6001 cttaaaaaaa ttacgccccg ccctgccact catcgcaata ctgttgtaat tcatttaaca
     6061 ttctgccgac atggaagcca tcacagacgg catgatgaac ctgaatcgcc agcggcatca
     6121 gcaccttgtc gccttgcgta taatatttgc ccatagtgaa aacgggggcg aagaagttgt
     6181 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga
     6241 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
     6301 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
     6361 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
     6421 tcaccagctc accgtctttc atcgccatac ggaactctgg atgagcattc atcaggcggg
     6481 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa
     6541 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     6601 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     6661 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     6721 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc
     6781 attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca ggatttattt
     6841 attctgcgaa gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg
     6901 ctgccaactt actgatttag tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt
     6961 tctatcagct gtccctcctg ttcagctact gacggggtgg tgcgtaacgg caaaagcacc
     7021 gccggacatc agcgctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg
     7081 tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat
     7141 gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
     7201 gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat
     7261 acttaacagg gaagtgagag ggccgcggca aagccgtt
//
