LOCUS       sequence-369420-        7190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Avs4 from Escherichia coli 382_h.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7190)
  AUTHORS   Gao LA, Wilkinson ME, Strecker J, Makarova KS, Macrae RK, Koonin EV,
            Zhang F
  TITLE     Prokaryotic innate immunity through pattern recognition of conserved
            viral proteins.
  JOURNAL   Science. 2022 Aug 12;377(6607):eabm4096. doi: 
            10.1126/science.abm4096. Epub 2022 Aug 12.
  PUBMED    35951700
REFERENCE   2  (bases 1 to 7190)
  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"; journalName: "Science."; 
            date: "2022-08-12"; volume: "377(6607):eabm4096. doi"; pages: " 
            10.1126/science.abm4096. Epub 2022 Aug 12"
COMMENT     Sequence Label: Ec2Avs4
FEATURES             Location/Qualifiers
     source          1..7190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1968..1979
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             4080..4088
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     terminator      4134..4181
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4401..5060)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4975..4994
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5061..5163)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5555..5572)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5689..6234)
                     /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(5769..5788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagcgcgtta attataagtg gagaaggcgg ctcaggaaaa acatctttaa taaaagagct
       61 ttatggcaat aaaggtcgga tggatgtctt ttatgtccat aaggcgacgg aattttctgt
      121 tagcaacttg aatgagtttt tgtcgggtgt ttttttaaag gattttattg aagcacatga
      181 aggagctgac aataaaacag ttgtaattga ttcagcggaa cacttaattg ctctagaaaa
      241 taccaatcct ataaaagagt acctatccag cttaattaaa tcaggatgga gagtgtggtt
      301 caccacgaga aatgcctatc tggatgattt gatttttcaa ttatatgaag tttacaatat
      361 tagttttaag tctgtacgcc tagaaaatct gaatgaagat acattattcg agcttgctaa
      421 aaaacataat tttgtcatcc ctgatgataa aaatctcaaa gttctcataa tgactccatt
      481 ttatctcaga gagtatttga gattttatca agaaaacaaa ggcgcaagct actttgaatt
      541 caaagagtct ttatggccaa gagtcattgc aaaacggtca ccacaacgag aacaattttt
      601 tatacgtctt gccgaagaaa gagctaattc tggaagattt ttcgtcattc cagacccgaa
      661 ctactcaaat gaaacagtag aaaaagcgtt ggtttccgat ggaataatta gctatgagcc
      721 agttcgtgga tattttatta cccacgatat ttatgaagaa tgggcactcg ataaattcgt
      781 cgagagcaac tttttatcct cagaaaatgc agaaatattt tttgatagaa tacaacaatc
      841 tttgcccata agaagagttt tcaggcggtg gctatcagaa aagcttaacg cggccaatga
      901 agaagttaac catcttattg tggaaacctt atcatctcgt aaaatatcta atttgtggaa
      961 agatgaagtt ctcgtttcca tgcttctatc cgactattcc gattactttt ttaaagttaa
     1021 taaagattca ctgttggaag acgacttgtg gctgctcaaa cggatatgct tgttgataag
     1081 aatagggtgc aaagagatag acaatagctt ttttgataga cttgatgtga gagcgccaga
     1141 tattttatca atggaatatg taataaccaa gcccaaagga aatggatgga aatcgcttat
     1201 caagttcatt aatgataatc tagaaaaaat aggtgtcaat aatcttaatt ttgtattgcc
     1261 tgttctctac gactggaact gtcacaacaa gtctggcgat tcaactaaat attccagcat
     1321 aatagcattg tctttttaca aatcggtaat tgaagatgat gtctatatta gcgatactga
     1381 tttttccaag agtttaatcc tcaccattct ttatggagca ggtgaaatta aaaatgaact
     1441 tgaaactatt attgacgaaa taatcataaa taattggaag caacataacg atccatatca
     1501 tttaatgagt gaattcattt taacaagaat ggaatgcttt aatgttgcca tggttattcc
     1561 ggaaaaagta attgctcttg caaaatgtct gtggatgtat gaatcaccag aaagcgatga
     1621 ttatttttat agttcaagat tagatataga tcatgaattt ggtattaata atggacacca
     1681 aaactattat ccagccagtg cttatcaaac ccccatatac gctctactgc aagctaattt
     1741 agaattaaca ttaaacttta ttattgaact aataaattac gcttcaaaaa aatatgtggc
     1801 atcaaacctt gataagggcc agattaaaac agcgaaactg ttactagatg atgggaagaa
     1861 aattgatctc cctatttcaa atcgtctttg gtgtatgtac cgaggtacac aagtaaatcc
     1921 tcatttatta gaatcaatat taatgtcact agaacgtttc tttcttgaaa gaggagaaaa
     1981 cacaaaagct caaacacttg aatattatct aaactatatc ctaactagag ctgaatcctc
     2041 tgcactcgtg gcagttgtcg caagcattgt atgtgcgttt catgagaaaa catttaatgt
     2101 tgctaaaatt ctattcagga caaaagagtt tttcatcttt gacacatctc gaatgatgca
     2161 ggatcaaaca cataagactc agcttacttt gctaaaaaac cctctcatca atcggataga
     2221 tgaacttcac gaaaatgaaa ggataagcac ctgtgacaaa aaacatcgga aactatcatt
     2281 agaaaacatt gcgcttcact accagttttt tagagcagag gaactaagcg aaagggaaac
     2341 agagaaaagg ctccaggaaa tttggaagat tcttgatgct cactatacag atttaccaga
     2401 taaagaacga gaaagtcatc aagataaaac gtggcgttta tatttggccc gtatggacaa
     2461 aagaaaaatg tcacctgaaa taaaggaaat agaaaattgt atagcaatag aattcaaccc
     2521 agagattgcc ccagagctta aggaatttag tgaaacatct ctgcgtgaag ctagcaaact
     2581 atttacacat tccggtttaa atatttgggc tgaaaacaga ttctataata gagataatta
     2641 caaaaaatat gaattatatg aaaacaatcc actgactgct ttgaacgagg caaaagaaat
     2701 atgggatcaa ctttcagaag gtaacatgag ggaggagttg ctgttttatc gagcgacgcc
     2761 atcgtatgta tgtgccgtgt tattgcggga ctttaaggaa atactagtaa aaaacgaact
     2821 ggaattttgt agggaagtta tatttgaata tgtaaatatc gttaacaatt atgattacat
     2881 gcaccagttc ggcgatggcc ttgttcctgc attatttgtg ctcccaatat taatagaaga
     2941 ttttccggac gaccgactaa ccatcaaact gctattaatc aacgcactaa tgaggcatga
     3001 tccactatcc atgatgggaa tggatagtat aaattcagta gccattcaag cagttcagta
     3061 cttgtggaat aatcaacctg agttcatgaa atctctgtat attggttatc tagttatctc
     3121 tcaaaaatat agggatgttc ggaatcgttt aaaacaagag gcatataagc agcataaata
     3181 tgaaattaac gacgatgact ttgagttgga gttatctaaa gaattagagt gtgtagtaca
     3241 gagtatcaaa gatgaaacta tcacggaatc cgcaatagac aatattgaat tcatagataa
     3301 ggatatatta ataactgttt ttcagcttgt accattgaat ggcgaacaca agaaacaagt
     3361 gccatttact gaggaaatta ttaggataat ttcacagcaa ttgctttcac aagacagaga
     3421 aggccgcatt gattatactt ttagatatca atttttaaag cactatgccc gttttatcct
     3481 tcatctacag gatagtgaaa agaacaaata ccttaagtac tttaccgata atttcacctt
     3541 aggtgaaggt atggcagatt tgttaaatga atttattagc gctgaagatt cattgaatat
     3601 gccagactca ttttggtatg tttggagtat atttaaaggt cacatcattg attcaataag
     3661 caatcaatgt tggaaatatg ataaagagcg cataatcgaa tcctttttat tttcaagagt
     3721 cccttggaag gatgaagcaa agacatggca tacattttca ccgggaaata gagcgttttt
     3781 tagtgattta agcgataaga taggcggtga accatctttt atatatagta tctgtaaact
     3841 cctttgtggc ataggaagcg aatttgttga cgatggaatt tattggatta gtaatgcaat
     3901 aaagaaccat gatatcgatt tatcacaaga taaatcaggg aatacactat tttacttgga
     3961 acgatatatg agaagatatc tattcaaaaa tcatgacaaa gttagacgga caccaaacct
     4021 aaaagcacgg tcaatgatgg ttctggattt tctagtcgag caatattcca ttacaggtta
     4081 cctccttaga gaagatatag cctagcgaaa atgcttgtca ctgttcgcaa taactagcat
     4141 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct caggcatttg
     4201 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
     4261 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt
     4321 tctgccattc atccgcttat tatcacttat tcaggcgtag caccaggcgt ttaagggcac
     4381 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcaata ctgttgtaat
     4441 tcatttaaca ttctgccgac atggaagcca tcacagacgg catgatgaac ctgaatcgcc
     4501 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatagtgaa aacgggggcg
     4561 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     4621 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     4681 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     4741 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     4801 ctatcccata tcaccagctc accgtctttc atcgccatac ggaactctgg atgagcattc
     4861 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     4921 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     4981 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     5041 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     5101 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
     5161 tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca
     5221 ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg
     5281 tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag gtgctccagt
     5341 ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg tgcgtaacgg
     5401 caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta tgttggcact
     5461 gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc
     5521 agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg
     5581 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
     5641 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
     5701 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     5761 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     5821 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
     5881 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
     5941 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
     6001 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
     6061 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
     6121 cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag
     6181 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
     6241 catcttatta atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta
     6301 gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc atgccccgcg
     6361 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     6421 cctaatgagt gagctaactt acattaattg cgttgcgcaa ggctgagaag gtctgatttg
     6481 aaccaaagcg agactttact tttttacgaa agcctcaaat agtcattgca aatgatatcg
     6541 tttgaatatc agttaatact taaaaaggat atagtaaaat tgattaagat agattgggaa
     6601 atatttaaag caaaattttc agcaaatcca caagacgctt ttgaatggat gtgttatctc
     6661 cttttttgca atgaatttcg agtgaaaact ggaattttcc gatataaaaa ccaatcggca
     6721 atagaaacca atccaatttc ctcaaatgga gtaaatgtgg gttggcaagc taaattttat
     6781 gaaaccgcat tatctggaca taaaaatgaa ttactagaaa cgcttataaa agcgaaacgt
     6841 gattaccctg atctgaataa aataattttt tacacgaatt cagagtgggg tcaaaacaag
     6901 ggaaaagaac cacaaggaaa aatagaagta gaaacgaagg caaaagcaaa caacttagaa
     6961 attgaatgga gatgctgtag ctttttcgaa tcgccgtttg ttgtcgatga ttgtaaccgg
     7021 atcgcttcgt acttcttcac tcgttctgat aatttattta gacttttgga gtcgtttgag
     7081 tctcataccc ttgcaattct ggacgatata gaaactacta taacctttgg cgatgaagat
     7141 gtttcaatca atcgttctaa tattatatct gaaattgaaa agtcagatac
//
