LOCUS       sequence-369873-        7979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon optimized EcAvs2 with C-terminal His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7979)
  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 7979)
  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-EcAvs2
FEATURES             Location/Qualifiers
     source          1..7979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        774..865
                     /gene="bla"
                     /label=AmpR promoter
     CDS             866..1657
                     /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(1828..1875)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1871..1889
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1921..1938)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    6443..6467
                     /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(6467..6486)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6468..6486)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6532..6609
                     /gene="lacI"
                     /label=lacI promoter
     CDS             6610..7692
                     /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(6629..6648)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tggtttgatc gaaggttaag cagttcccca actgacttaa ccttcgaaaa aaccacctgg
       61 ggaggtggtt tgatcgaagg ttaagtcagt tggggaactg cttaaccgtg gtaactggct
      121 ttcgcagagc acagcaacca aatctgtcct tccagtgtag ccggactttg gcgcacactt
      181 caagagcaac cgcgtgttta gctaaacaaa tcctctgcga actcccagtt accaatggct
      241 gctgccagtg gcgttttacc gtgcttttcc gggttggact caagtgaaca gttaccggat
      301 aaggcgcagc agtcgggctg aacggggagt tcttgcttac agcccagctt ggagcgaacg
      361 acctacaccg agccgagata ccagtgtgtg agctatgaga aagcgccaca cttcccgtaa
      421 gggagaaagg cggaacaggt atccggtaaa cggcagggtc ggaacaggag agcgcaagag
      481 ggagcgaccc gccggaaacg gtggggatct ttaagtcctg tcgggtttcg cccgtactgt
      541 cagattcatg gttgagcctc acggctccca cagatgcacc ggaaaagcgt ctgtttatgt
      601 gaactctggc aggagggcgg agcctatgga aaaacgccac cggcgcggcc ctgctgtttt
      661 gcctcacatg ttagtcccct gcttatccac ggaatctgtg ggtaactttg tatgtgtccg
      721 cagcgcccgc cgcagtctca cgcccggagc gtagcgaccg agtgagctag ctatttgttt
      781 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      841 tcaataatat tgaaaaagga agagtatgag ggaagcggtg atcgccgaag tatcgactca
      901 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
      961 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     1021 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     1081 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     1141 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     1201 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     1261 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     1321 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     1381 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     1441 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     1501 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     1561 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     1621 cgtgaaaggc gagatcacca aggtagtcgg caaataatgt ctaacaattc gttcaagccg
     1681 aggggccgca agatccggcc acgatgaccc ggtcgtcggt tcagggcagg gtcgttaaat
     1741 agccgcttat gtctattgct ggtttaccgg tttattgact accggaagca gtgtgaccgt
     1801 gtgcttctca aatgcctgag gtttcagcaa aaaacccctc aagacccgtt tagaggcccc
     1861 aaggggttat gctagttatt gctcagcggt ggcagcagcc taggttaatt agccgctgct
     1921 gtggtgatga tggtgatggc ttccgccacg ggtgaagctg gttttgtggc taccggtcag
     1981 ctggctgtcc tcatagtaaa acgcaccgct gaattcttga cggccatagt ggtggctgct
     2041 aatcaggctg ccaccgataa cacctttctc gcccagcagg gtccacacga tttcaaaacc
     2101 gttatcacgc agcatggtca ggaacggctc tttcttaacc agcaggtgcg gcttgctcgc
     2161 ataaaccgcc tccgccgcaa agcacaccat ggtgccgttt tcgtcgttga agctaccctc
     2221 cacttcgccg cttttcagac ccatcttctc aaaaatcagg ttgctcggtt tcaggaagtt
     2281 cagggtttcg attttgctct tatcgaactc ttcctcccac aggtagttaa tgctggtaac
     2341 gctcacgtcc gcgatcttcg caccgctctc acgatcggta acgctggtcc agtcgctgcc
     2401 accatagtaa tcgcttttaa agctcttgaa cgcctcgctc caatagtatt cacggttgaa
     2461 cagctggtaa cgatcggtgc actccggcat ccaacggccc ataaagtctt gcgcgatcgc
     2521 ccagcaacga aagttttcga attcctcaac tttcacaata tagctacgga tctggtacca
     2581 cacctcttta cgcgggtgac cccaatcgtc gttgccaatg attttcggtt ccttccagct
     2641 cgggtggctc tccagaacga tccattcatc accgttatcg tctttcacct caatgaacgc
     2701 gtagctgttg gtgatggtgg tgctgctctt aacccagtct tcgttgctgc aggtccagtc
     2761 aaacacctca tcgttcagcc accacatttc tttgttgctg attttcttgg taccggtctc
     2821 cttcagcaga atggtcggat cgatgtcacg aacatacggt tcccacgggc cctggtacgg
     2881 gttttcttta cgctcatcac cgtagccctc aaaacgggtg aagttgtccg ccagacgcgc
     2941 catatattca tagtacgcaa tccactggta tttcttaccg atacgctctt gaaacgcttc
     3001 acgacgacca cggccggtac cgatctgttg atcgaagtta ccgtgtttct ccgggtccca
     3061 gcccagttca atcacacggt taaagatgaa ctgttgcgcc agacgcaggt cgaagtgctt
     3121 atcggtctcc agcaggttgt tgttgtgatc cagatacggc tcaatttcgt tctcaaattc
     3181 gctcagcagg tcgtagctca ggctgttctt gaacagtttc ttgctcgctt tgatctcttc
     3241 ctgcgcctta cggccaacca cacgaccaaa tttaatgccc tcgcacttat cgtcatagat
     3301 gaacgggtcg gtcgcatcgt acaggtcttt ctgttggtcg gtcagtttgc acttaaaggt
     3361 tttgaaaact tgcttacgat ccaccggggt ttcgttgaac ttgcaaccgc tccagtcgct
     3421 gtggttatag ttggtaccaa tggtgtaacg gctaaaatcg ccgtcctcca tgatgctgct
     3481 ccacagctca cgatacggtt ctttatcgta caggctttcc agttcctctt tgctcggaat
     3541 cttgtccggc cagatgctgt tatacggcgg acgggtcttg ctcagctcaa tgctctccag
     3601 ttccaggccc aggtggttcg cgaattcaat gatctcacgc gcgtaatcac gcagcaggat
     3661 gttcggataa acgtatttgc tgcagaacac cttctgatac acggtctccg ccaggtattt
     3721 cagttcctta atgttgtcgg tacgcagggt gcaacccagc gccaccgcaa agatacgttc
     3781 ccaaacatac ggatcgttca cgccgtagaa tttgtcaatg atcttacgca gaaccgggat
     3841 acgcggctcc agcaggctaa ccagcgcttt ggtgctgcaa tcacgcagtt cacggttgct
     3901 gctggtcagg aaccagcaca ggctggtcgc aaccagttca atgctctcgt cgctcacgaa
     3961 gctcttatcg gtacgcgccc acgcccaatc gatcaggtga cgaaacgcgc tgtcctcgct
     4021 atatttgtac ttcagttcgg tggtccagaa gctgtcacgg ttcgccaggc tgtagtcctt
     4081 cagccaatcg tgcagaaagt tcgcgttgaa cgggtgacca accaggccgc taatgctgat
     4141 caccgcctcc agaaagtgat cgaagctgtc cttgaacttg aaaacgtgct cgttgatgaa
     4201 cggacggatt ttttcgaagt caatcgcctt gatgtcacgc caaatcaggc tgtcgatgaa
     4261 cgcttccagc agtttcaggt tgttgctaaa ctccggcagg aattcataca gctccttttc
     4321 gtaacgctcc ggcagctgga tgctcagcgc ttccacaata ccgcttttga tataaaaatc
     4381 gcactcgtcg tggaagtatt tcttcagacg gccatccggc ttaaactcgc tttcgatgtt
     4441 ctcaacgtcg ttcagcagga agttcacggt caggtgatcg tcaaaacgtt cgaacgcaac
     4501 ataaaccact tcctcggtgc tgttatcgtc gttacgcaca atacctttgg tcagcaggcc
     4561 ttcatcgatc agcgcgctca ggaaggtctt gtccgcaacg taatcgttaa ccacgctttg
     4621 aaccacgctg tgcgcgtctt tcaggctaat gctgttacga ccgatctcca gtttaaactt
     4681 aatgatttcg ttcagcgcat ctttcaccag cggaaagctc gggtcgaacg cgtatttctt
     4741 cgggctcgcc aggctcttgt taacgccctc caccagaaag ttgaaaatgt tgctgatacc
     4801 gttgaagcca accggcacct tggtcagacc gtttttctta atgccttcgc acagcagttt
     4861 cagaaacagc gggttcttga actccgggtt caggttcggg ctgctcggac gttcgatgtt
     4921 atagtagtca taaaacaggc taaccgcctc cagttccacg ttctgaaaac caatgtgttc
     4981 gtggatctcg aagttgttac gaaccacgtt ctcgtggcta atggtaacgt tacggtaggt
     5041 ggtacgcacg ctcatgatca ggcccagcca ctcaaagcaa cgaatttcat caacgaagct
     5101 gttgatgttg tcgttccaaa atttgttgcc gttaccctcg ttaatcgcgt cgatgaaaac
     5161 cagcacgcgt ttaccggttt tcttgccata caggttcagc ttttccagaa actcacggct
     5221 ggtaattttc agctgcagac gcttgaagat ttggctccac gggctctcat cgctggtcag
     5281 ctgttgaccc agaatcagca ggctcgggta gccgctcgca atacggtttt tgataacgtc
     5341 cgccagcagg tggcttttgc cgatacccgc cttgccttcc agcagcagga acgggttgtt
     5401 cgccagcttc acggtggtgc tgttaataaa gatacgcagc tcgttgcacg cgtaatcgaa
     5461 ttcacgcagc atacgcaggg tgctgctata tttgtcgctg tagtgcttgg tctcgcccac
     5521 ttgttcgctt ttctcacgca gttcccacag aatgctaacc gcgccaccga tcgcggtttg
     5581 gcagctgctc acatagttgt tgaacttgtt gatcgggatt tcatccagct tgctgaagtt
     5641 gatctcctgg taaatgtcgc tgatttcctt caggctttgg ttaatctcga acagttcgct
     5701 gctgataaca tcgcagctgt gcagtttctt acccgccacc agaaagctgt caatgtggct
     5761 ataaaagatt ttgctgaaat cgttggtacg gcccagaccg tcgaaaatct ccgcgatttc
     5821 cagcttaacg ttcagctccg gggtgtaacg gccacccagg tccgcaatcg cctggctgtt
     5881 cttctcgttc agccaatcgt cgctcaggtg gaaggtcagt ttttcatcaa tgctacgcag
     5941 gatcagcttc acttggatca gttcctcgtt aatgtcgaag atcttttcct tgtaggtttc
     6001 ctcaatgtgc agtttcttca gtttgctgca gttgttgatc ttcaggctca gcatctcata
     6061 gaacaggctc agctgttgct gggtcggcgg ggtgatgctc gggtattctt taaacttgtc
     6121 cagcagaatc tctttatccg ggaattcctt aaagaagatg tgctcgttca gcatttcaaa
     6181 cagcggctcg ctgtgataga acgggatcgc accgctctca accgggtagg tttcctcgaa
     6241 ttcaatcgcg gtttcctcga tcagctgata cagacgatcc acgtaacgtt tttgcggaaa
     6301 cagcgctttc ttgatgcagc cataaccttc ctggctaatg aattggtcga tcagcttggt
     6361 cgcaacgtag gtcgccacgg taatgctgat cggttccatg gtatatctcc ttattaaagt
     6421 taaacaaaat tatttctaca ggggaattgt tatccgctca caattcccct atagtgagtc
     6481 gtattaattt cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc
     6541 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     6601 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     6661 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     6721 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     6781 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     6841 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     6901 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     6961 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     7021 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     7081 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     7141 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     7201 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
     7261 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
     7321 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     7381 accgagtccg ggctgcgcgt tggtgcggac atctcggtag tgggatacga cgataccgaa
     7441 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     7501 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     7561 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     7621 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     7681 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc
     7741 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     7801 gactaacatg agaattacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa
     7861 gagataaatt gcactgaaat ctagagcggt tcagtagaaa agatcaaagg atcttcttga
     7921 gatccttttt ttctgcgcgt aatcttttgc cctgtaaacg aaaaaaccac ctggggagg
//
