LOCUS       sequence-369864-        8288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon optimized EcAvs4 with C-terminal His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8288)
  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 8288)
  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: pCDF-EcAvs4
FEATURES             Location/Qualifiers
     source          1..8288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1002..1093
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1094..1885
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      complement(2056..2103)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2099..2117
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2149..2166)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    6980..7004
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(7004..7023)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7005..7023)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7069..7146
                     /gene="lacI"
                     /label=lacI promoter
     CDS             7147..8229
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(7166..7185)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actaacatga
       61 gaattacaac ttatatcgta tggggctgac ttcaggtgct acatttgaag agataaattg
      121 cactgaaatc tagagcggtt cagtagaaaa gatcaaagga tcttcttgag atcctttttt
      181 tctgcgcgta atcttttgcc ctgtaaacga aaaaaccacc tggggaggtg gtttgatcga
      241 aggttaagca gttccccaac tgacttaacc ttcgaaaaaa ccacctgggg aggtggtttg
      301 atcgaaggtt aagtcagttg gggaactgct taaccgtggt aactggcttt cgcagagcac
      361 agcaaccaaa tctgtccttc cagtgtagcc ggactttggc gcacacttca agagcaaccg
      421 cgtgtttagc taaacaaatc ctctgcgaac tcccagttac caatggctgc tgccagtggc
      481 gttttaccgt gcttttccgg gttggactca agtgaacagt taccggataa ggcgcagcag
      541 tcgggctgaa cggggagttc ttgcttacag cccagcttgg agcgaacgac ctacaccgag
      601 ccgagatacc agtgtgtgag ctatgagaaa gcgccacact tcccgtaagg gagaaaggcg
      661 gaacaggtat ccggtaaacg gcagggtcgg aacaggagag cgcaagaggg agcgacccgc
      721 cggaaacggt ggggatcttt aagtcctgtc gggtttcgcc cgtactgtca gattcatggt
      781 tgagcctcac ggctcccaca gatgcaccgg aaaagcgtct gtttatgtga actctggcag
      841 gagggcggag cctatggaaa aacgccaccg gcgcggccct gctgttttgc ctcacatgtt
      901 agtcccctgc ttatccacgg aatctgtggg taactttgta tgtgtccgca gcgcccgccg
      961 cagtctcacg cccggagcgt agcgaccgag tgagctagct atttgtttat ttttctaaat
     1021 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1081 aaaaaggaag agtatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     1141 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     1201 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
     1261 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     1321 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     1381 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
     1441 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
     1501 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
     1561 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
     1621 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     1681 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     1741 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     1801 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga
     1861 gatcaccaag gtagtcggca aataatgtct aacaattcgt tcaagccgag gggccgcaag
     1921 atccggccac gatgacccgg tcgtcggttc agggcagggt cgttaaatag ccgcttatgt
     1981 ctattgctgg tttaccggtt tattgactac cggaagcagt gtgaccgtgt gcttctcaaa
     2041 tgcctgaggt ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc
     2101 tagttattgc tcagcggtgg cagcagccta ggttaattag ccgctgctgt ggtgatgatg
     2161 gtgatggctt ccgcccagaa tgttctcacg gctcatatag ccaaccacgc tacctttttc
     2221 aaccaggaag tccaggatca ccagaacctc ttgtttcagc tgcttggtac gacgaatacg
     2281 ttcacgctcg ttgttgatgt aacgacgcac caggcattcc agatagtaaa cggtatcgtt
     2341 ttccagtttc ttctcccaca gtttcttgtt cacgctcaga atctcgctca gccaggtgat
     2401 accctggttc aggtagcagc tcgcgatgtt gttcaggctc ttcgccaggc tatacagggt
     2461 gctcgggcag tgacccatgg tacggctaac atcgcaaaag aattggctgt tgctgtcttt
     2521 aaaggtgtgc cagccgttgc tgttttcctt ccacgggctc tccgcgaaca ggtagctttt
     2581 aatgatcttg tcaacgtacc agtaacggtc gccatctttg cacagggtaa ccaccttgtc
     2641 gaagaacagg tcccaaactt tccagaactt cgcataggtg ttcagacggt cctccaccag
     2701 aataaatttc ttgaacagtt cgctgatcgg ctcgctgccg ttaaaaccat ccaggaacgg
     2761 tttaatgtag tccgggatgt cgctaaccgg cgcatgcagg gtaaagtacg cgaaacgttc
     2821 cagaaagctc tgacgcagcg catagtccac acgatcctca cgaacgctgc tcagcaggct
     2881 ggtgctgaaa cgcttcacga tcaggctaac cagtttcttg tgctcaatgt tcacggtgtc
     2941 aaccgggatc agttgaaacg cggtgttcag aatgtgcagg tcaactttat ccatgctacc
     3001 cagatcgtca atgctgatct tgttatcaat aacgttgctg atgcagtgtt tatagttgtt
     3061 caggaacacc ttgttgatgt tgattttctt gatgtcgtaa tcaacctgac ggtagctttc
     3121 gtgaatgatt ttacggctca gaatcacgta tttcggcttc aggatcagaa aaccgaacag
     3181 caggctttgc atatcgtcaa agtagtccag ccacagcttg tgaatcacca tgctcgggaa
     3241 aacgctataa cggccacccg ccatgccgat gctgtgatcg ttgaacaggg tcagcagcag
     3301 aatggttttg atggtttcac gctccatcgg gtagttgtga taaataaccg gcagcgcgct
     3361 gatcgcgctg gtggtaccat cttgcacctg atagttgtaa ccttctttca gcggcagctt
     3421 gctgtacgcc agaacaatat ccttgcaata ctcacgctcc tcgttgttca gctggttaaa
     3481 gtaatccagc agcagcacgc tgcaaacgtc cgccgggatg ttaccgttca gcagacggaa
     3541 gtcttcgcca ccttcctcgt tcagtttctt aatgatttct ttggtctctt gcagcgcaat
     3601 ctgcgggtta tcctcgtaca tgccatagtt cttgtaacgt tcgtctttct cacgcttgta
     3661 gctcgcccac agcttcaggg tcacgtactt catgtgttcg ctgtttttct taatcgcttc
     3721 ctcgctgtac tgtttcagct tcgggtcgat ctccgggttg aagctaattt cgataccctc
     3781 atctttttcc ttggtggtga ttttcatctt acgacggtcc atacgcgcca ggcacagacg
     3841 ccaggtttta tccgcttcgg tctcttgcgc ttcgtccggc agctggttat agtatttgtc
     3901 aaaaatatcc cacagcactt gctgacgctc gatcgcgtcc ttttcggtaa cgttctcgct
     3961 acggaagatc tggtagtgca gcgccaggtt ctccagatag gtgttacggt gaacatcgtc
     4021 gcacgcttta atacggtctt cctcgtgcag gctgttacgg taatcggtgc caccgaagcc
     4081 gtcacgcagg ctgatcaggc tgctcttgtg ggtacgatcc agcaccatac ggttcatatc
     4141 aaaacgaaag aagtctttgg tttggaacag caccttcgca acgttaaagg ttttttccgg
     4201 gaacgccaga acaatgctgg taaccaccgc gctgatgctc gcgcttttgg tgttacgcag
     4261 cagaaacagc agccagctct ccagcacctt gctgtccgcg tttttgaagt tttccagaaa
     4321 gaacttctcc agcgccatgt ggatgctgct cagcaggtag gtgctaactt gggtaccacg
     4381 atagctgcac cacagacggt tgcaaatgta ctgtttgata aatttgcctt cctcaatgaa
     4441 cacatcaact tcctcgatct cgtttttcgc aaagtggctg tgcgcgaagc agatggtggt
     4501 cttgttggta aagtccagaa tgaaatcgat ggttttcttg aattggctct gcagcagcca
     4561 atagatcggg gtctggtacg ggctttccgg atagtagtcg tggtggctac gaaacagacc
     4621 gaattcgtcc tcaatatcca tgctgtggta acgctcgccg gtttctttca gcggacgata
     4681 ccaaaacagg tccgccagtt gcagcacgta ctccggcagg ctcgcccaaa ccggatagct
     4741 gtccagatcg gtcaggatca gggtcatcag atcgaaatac ggggtaccgt gttccttcca
     4801 acggtttttc agcaccttaa ccagcacttc ctccatctcc ggtttgatca tcgccgcacc
     4861 gtgcagaatg gtgtgcagga tgttcttttc gttatcacgg ccgctcaggt acacgtcctc
     4921 atcaatggtc cactgatagt atttcagcgc gatcaggctg ctcagacggg tggtttcacc
     4981 cactttgtta cgttggttcc actcctggat aaccggcaga ataaagttga tgttacgaat
     5041 gccgatttcg tccaggttct cataaatgaa ttggatcacg ctctgccagc cggtaccttt
     5101 cggcttggtc agaacgtact tgatgctcag cagatcgctg ttgctcacgc ccagttgttt
     5161 cagcaggtca taatcaacgt ccttgcacgc cagacgcagc agaaaggtca gacgtttcag
     5221 caggttctgg tcgctgctca gcagataacg cttgaagtag ttaaagaaaa tgctgctgtt
     5281 atcgctcagc agaaccgcca cccacagctc gtctttccaa aagttgctga tgccttcacc
     5341 gcaaacaatc tccgcgatga acggcttaat gctctgatcg tccagcagca gacgttcgct
     5401 gatccagtta cgaaagctac gacgaaccgg caggctctcg ccgattttct caaagaattc
     5461 gttgttgttc gccttacgaa tgtagtccac gctgattttc ttttccagcg cccattcctc
     5521 ataaatatcg tgggtgatga aatagcccgc cgcttcgtag cccacaatac catctttaac
     5581 cagctcgtcc aggataccgg tatcgcacgc cgggctcaca aagaattggc cttcgctcgc
     5641 acgctgaaac gcggtcgcca ggaagcactg ctcacggctc ggtttgttct tcacgatgat
     5701 cttgttccac agcttttcct tgaagctaac gtagtcaatg ctttcaccgg tataaaaacg
     5761 caggtactcg ctcagataga acgggttctt gatcagctcc agcagacgaa catcttgcgg
     5821 caggctgaaa ccgttgttgt cgctcagctc gatcagttcg ccacgctcca ggtttttaat
     5881 caccaggtta cccggggtga tcttgtaaat atcgataaac gcatagttca ggtccgccag
     5941 gtagttgtta cgggtggtga aaaccacctg ccatttatcc ttaatcagaa cggtcagaaa
     6001 ttccttgaac gggtcgatgt tggtcagttc cagcagtttc tccgcgctat caaccacgat
     6061 caccttacgc agctcgtctt ggtgcgcgtt gctaaaatcg tccaggccgt gcgcaccgaa
     6121 cagttcgtta atgctgtctt tcttgaactc gctcgcttta aaaacgtaga acggggtata
     6181 ctgcttttcc gcctcgtaaa ttttcttgat caccgcggtt ttgccaacgc caccctcgcc
     6241 gctaacaatc acgattttct tttgaaccag gttttcgtgc agcagctcga tgcagtgacg
     6301 acgatcaatt tcgatgctac ggtccttgaa ctcaatgttg gtctggattt cgtacagcac
     6361 gttctcggtg tgtttacgct tttcctccag cagatcaaag atgctttcgt tcaggctaaa
     6421 gaagtgtttc gcaatctttt cgttctcaac gatcacgaac gggctctcaa agaagctcgc
     6481 cacacgccaa acaatttcga taccgctctc atacgccttc tgatccactt caattttgat
     6541 cttcgggtcg ttgctgttac caacggtctc caggtagttg tccgcgtttt tatcgccttc
     6601 cggctcgtgg ctcttacggc cttgacccca ctcctggttg gtgtagaaaa tgattttgct
     6661 cagacccgga tacgctttct tgctcttttc aatcatctcg atcaggtccg ccttgttatc
     6721 gctcagtttg gtatcgtaaa acttgctttg ccagccaatg atttcgttgt ctttggtaat
     6781 cgggttggtc tcgatgccgc tctggttttt gtaacggaag atacccgccg gcatcttaaa
     6841 ttcttggcag aacagcagat agcaaaacca ttcgaagtta ccctgcgggt tctcgctaaa
     6901 tttcgccttg aagttgtccc agttcggctt caccatggta tatctcctta ttaaagttaa
     6961 acaaaattat ttctacaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta
     7021 ttaatttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa
     7081 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     7141 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     7201 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     7261 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     7321 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     7381 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     7441 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     7501 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     7561 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     7621 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     7681 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     7741 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     7801 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     7861 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     7921 gagtccgggc tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga taccgaagac
     7981 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     8041 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     8101 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     8161 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     8221 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg
     8281 accgatgc
//
