LOCUS       sequence-304755-        4398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the retron-TIR defense system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4398)
  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 4398)
  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: pLG005 (Retron-TIR)
FEATURES             Location/Qualifiers
     source          1..4398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2178..2225
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2445..3104)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3019..3038
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3105..3207)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3599..3616)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3733..4278)
                     /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(3813..3832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
       61 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgcaagtta atgactattg
      121 tgagcgagaa acgcgctact actatatata gacagacaag atgcacttac tgaataaata
      181 ctcataacgg agaaaccagc tgtatagtga acaatagatt tccagtagca tatttttact
      241 tcacttttag ttattaatat gataatcata aactacggct ctgccttaaa tttgtgaggt
      301 tgtttcgcct cgaaggaact aatgttagga catacgccac cgttcagtcg atggtaacgc
      361 ttcttaacta gtggtccgct aagtgatgcg caaagtgatt gggcagagcc gaaacgttta
      421 caatccgata ggagttggtt ttgtcgctac atgataaatt attaatgcat aacttcgcat
      481 tagccaataa aaaaagccct gacttcatat ctgaacttcc tcaaattgaa cctaaaccat
      541 acagcaatgg acataaaatt aaatggataa accacacact tactagcact gaagttactc
      601 cccctgataa cctgattaaa atatgcatat tgattgagtc aggggaaatt gctataacat
      661 cagtaagtga tattgccaat ttacttggag ttcctgctgg ccaattactt tatatactat
      721 atcgtaaaaa agataattat cgtacttttg aaatagaaaa gaagaatggt aaaaaaagag
      781 tcattaatgc tccttgtggc ggtctatcga tactccaaac gagactaaag cccgttcttg
      841 aatatttcta caggccaaag aaatctgctc atggttttat aaaaggaaag agcatcatta
      901 ctaatgctgg gatgcatatt aaaaaaaatt ttgtcgtaaa cattgatcta gaaaactatt
      961 tcgaatcaat aagttttgct agggtttatg gaatatttaa aagtaaacct tttaattttg
     1021 ctcatcctgc agctactgtt ttagctcagt tatgtactca caatggaaaa ttacctcaag
     1081 gtgcgtgtac atcgccaata ttagcaaata ttgcatcagc ttctctagac aaacagctca
     1141 cccaatttgc aggaagaaaa aaaatatctt attctaggta tgctgacgac ataacttttt
     1201 ctttcaatca gagaaatatt gatataatca aaaaaaacga cgacggaagt tatagtctta
     1261 gtgaaactat agacaatatt atttcaaaaa atggctttaa aataaattat gataaattta
     1321 gagttcaaac cagaaataca agacaaagtg ttactggctt agtggttaat gataaagtta
     1381 acattaacag aagatatata agaattacac gttcaatgat tcatagatgg acagatgata
     1441 agctaaagta tgcacttctc tttgctacag aaaaaggata tcaggcaaag gataataacc
     1501 acgcaattca aattttccga aatcatattt atggaaggct tagctttata aaaatggtta
     1561 gagggaaaga ctatccagga tatttaaaac tgatgtcata catgagtcat aacgatccat
     1621 taaaaaccca agaaggattg cgagcaatga aagaaacaga aaactttgat gtttttatat
     1681 gccatgcaag cgaagacaaa aaagacattg caattccaat atatgacgag ttaactaaac
     1741 ttaaaatttc agccttcata gatcatgttg agataaaatg gggcgactcc ttaattgata
     1801 aaataaatgc agcactagtt aaatcaaaat atgtcatcgc tattttatct gctaattcag
     1861 tcaataagga atggcctcaa aaagaattaa gagcagtttt agccagcgaa atatcgagtg
     1921 gcgacgtaaa acttttgacc ttattaaaaa aagaagacga ggaggtcgta aacctatcat
     1981 tacctttact tagtgataag ttttatatgg tctatgataa taatcctgaa gtagtcgcca
     2041 acaatattaa atcactctta caacgataat tctctcacaa aagaaaatgt gcagattgat
     2101 gcgtattaag tattaatctg cacatacaaa aaaaataata aaataataca tttttcataa
     2161 cttgtaggtc gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     2221 ttttgctgaa acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
     2281 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
     2341 ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc
     2401 gtagcaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc
     2461 cactcatcgc aatactgttg taattcattt aacattctgc cgacatggaa gccatcacag
     2521 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     2581 ttgcccatag tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     2641 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     2701 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     2761 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     2821 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcatcgcc
     2881 atacggaact ctggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     2941 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     3001 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     3061 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     3121 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     3181 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc
     3241 ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca
     3301 ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg
     3361 atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct cctgttcagc
     3421 tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct agcggagtgt
     3481 atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga
     3541 gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct
     3601 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
     3661 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
     3721 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
     3781 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg
     3841 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     3901 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     3961 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     4021 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     4081 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     4141 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     4201 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     4261 agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt ctagatttca
     4321 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa
     4381 ttctcatgtt agtcatgc
//
