LOCUS       sequence-366510-        8223 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 8223)
  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 8223)
  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..8223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             4858..4875
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4907..4925)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4921..4968
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5139..5930)
                     /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"
     promoter        complement(5931..6022)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6070..6808)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(7018..8100)
                     /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     8062..8081
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8101..8178)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg gtgaagccga actgggacaa cttcaaggcg
      121 aaatttagcg agaacccgca gggtaacttc gaatggtttt gctatctgct gttctgccaa
      181 gaatttaaga tgccggcggg tatcttccgt tacaaaaacc agagcggcat cgagaccaac
      241 ccgattacca aagacaacga aatcattggc tggcaaagca agttttacga taccaaactg
      301 agcgataaca aggcggacct gatcgagatg attgaaaaga gcaagaaagc gtatccgggt
      361 ctgagcaaaa tcattttcta caccaaccag gagtggggtc aaggccgtaa gagccacgag
      421 ccggaaggcg ataaaaacgc ggacaactac ctggagaccg ttggtaacag caacgacccg
      481 aagatcaaaa ttgaagtgga tcagaaggcg tatgagagcg gtatcgaaat tgtttggcgt
      541 gtggcgagct tctttgagag cccgttcgtg atcgttgaga acgaaaagat tgcgaaacac
      601 ttctttagcc tgaacgaaag catctttgat ctgctggagg aaaagcgtaa acacaccgag
      661 aacgtgctgt acgaaatcca gaccaacatt gagttcaagg accgtagcat cgaaattgat
      721 cgtcgtcact gcatcgagct gctgcacgaa aacctggttc aaaagaaaat cgtgattgtt
      781 agcggcgagg gtggcgttgg caaaaccgcg gtgatcaaga aaatttacga ggcggaaaag
      841 cagtataccc cgttctacgt ttttaaagcg agcgagttca agaaagacag cattaacgaa
      901 ctgttcggtg cgcacggcct ggacgatttt agcaacgcgc accaagacga gctgcgtaag
      961 gtgatcgtgg ttgatagcgc ggagaaactg ctggaactga ccaacatcga cccgttcaag
     1021 gaatttctga ccgttctgat taaggataaa tggcaggtgg ttttcaccac ccgtaacaac
     1081 tacctggcgg acctgaacta tgcgtttatc gatatttaca agatcacccc gggtaacctg
     1141 gtgattaaaa acctggagcg tggcgaactg atcgagctga gcgacaacaa cggtttcagc
     1201 ctgccgcaag atgttcgtct gctggagctg atcaagaacc cgttctatct gagcgagtac
     1261 ctgcgttttt ataccggtga aagcattgac tacgttagct tcaaggaaaa gctgtggaac
     1321 aagatcatcg tgaagaacaa accgagccgt gagcagtgct tcctggcgac cgcgtttcag
     1381 cgtgcgagcg aaggccaatt ctttgtgagc ccggcgtgcg ataccggtat cctggacgag
     1441 ctggttaaag atggtattgt gggctacgaa gcggcgggct atttcatcac ccacgatatt
     1501 tatgaggaat gggcgctgga aaagaaaatc agcgtggact acattcgtaa ggcgaacaac
     1561 aacgaattct ttgagaaaat cggcgagagc ctgccggttc gtcgtagctt tcgtaactgg
     1621 atcagcgaac gtctgctgct ggacgatcag agcattaagc cgttcatcgc ggagattgtt
     1681 tgcggtgaag gcatcagcaa cttttggaaa gacgagctgt gggtggcggt tctgctgagc
     1741 gataacagca gcattttctt taactacttc aagcgttatc tgctgagcag cgaccagaac
     1801 ctgctgaaac gtctgacctt tctgctgcgt ctggcgtgca aggacgttga ttatgacctg
     1861 ctgaaacaac tgggcgtgag caacagcgat ctgctgagca tcaagtacgt tctgaccaag
     1921 ccgaaaggta ccggctggca gagcgtgatc caattcattt atgagaacct ggacgaaatc
     1981 ggcattcgta acatcaactt tattctgccg gttatccagg agtggaacca acgtaacaaa
     2041 gtgggtgaaa ccacccgtct gagcagcctg atcgcgctga aatactatca gtggaccatt
     2101 gatgaggacg tgtacctgag cggccgtgat aacgaaaaga acatcctgca caccattctg
     2161 cacggtgcgg cgatgatcaa accggagatg gaggaagtgc tggttaaggt gctgaaaaac
     2221 cgttggaagg aacacggtac cccgtatttc gatctgatga ccctgatcct gaccgatctg
     2281 gacagctatc cggtttgggc gagcctgccg gagtacgtgc tgcaactggc ggacctgttt
     2341 tggtatcgtc cgctgaaaga aaccggcgag cgttaccaca gcatggatat tgaggacgaa
     2401 ttcggtctgt ttcgtagcca ccacgactac tatccggaaa gcccgtacca gaccccgatc
     2461 tattggctgc tgcagagcca attcaagaaa accatcgatt tcattctgga ctttaccaac
     2521 aagaccacca tctgcttcgc gcacagccac tttgcgaaaa acgagatcga ggaagttgat
     2581 gtgttcattg aggaaggcaa atttatcaaa cagtacattt gcaaccgtct gtggtgcagc
     2641 tatcgtggta cccaagttag cacctacctg ctgagcagca tccacatggc gctggagaag
     2701 ttctttctgg aaaacttcaa aaacgcggac agcaaggtgc tggagagctg gctgctgttt
     2761 ctgctgcgta acaccaaaag cgcgagcatc agcgcggtgg ttaccagcat tgttctggcg
     2821 ttcccggaaa aaacctttaa cgttgcgaag gtgctgttcc aaaccaaaga cttctttcgt
     2881 tttgatatga accgtatggt gctggatcgt acccacaaga gcagcctgat cagcctgcgt
     2941 gacggcttcg gtggcaccga ttaccgtaac agcctgcacg aggaagaccg tattaaagcg
     3001 tgcgacgatg ttcaccgtaa cacctatctg gagaacctgg cgctgcacta ccagatcttc
     3061 cgtagcgaga acgttaccga aaaggacgcg atcgagcgtc agcaagtgct gtgggatatt
     3121 tttgacaaat actataacca gctgccggac gaagcgcaag agaccgaagc ggataaaacc
     3181 tggcgtctgt gcctggcgcg tatggaccgt cgtaagatga aaatcaccac caaggaaaaa
     3241 gatgagggta tcgaaattag cttcaacccg gagatcgacc cgaagctgaa acagtacagc
     3301 gaggaagcga ttaagaaaaa cagcgaacac atgaagtacg tgaccctgaa gctgtgggcg
     3361 agctacaagc gtgagaaaga cgaacgttac aagaactatg gcatgtacga ggataacccg
     3421 cagattgcgc tgcaagagac caaagaaatc attaagaaac tgaacgagga aggtggcgaa
     3481 gacttccgtc tgctgaacgg taacatcccg gcggacgttt gcagcgtgct gctgctggat
     3541 tactttaacc agctgaacaa cgaggagcgt gagtattgca aggatattgt tctggcgtac
     3601 agcaagctgc cgctgaaaga aggttacaac tatcaggtgc aagatggtac caccagcgcg
     3661 atcagcgcgc tgccggttat ttatcacaac tacccgatgg agcgtgaaac catcaaaacc
     3721 attctgctgc tgaccctgtt caacgatcac agcatcggca tggcgggtgg ccgttatagc
     3781 gttttcccga gcatggtgat tcacaagctg tggctggact actttgacga tatgcaaagc
     3841 ctgctgttcg gttttctgat cctgaagccg aaatacgtga ttctgagccg taaaatcatt
     3901 cacgaaagct accgtcaggt tgattacgac atcaagaaaa tcaacatcaa caaggtgttc
     3961 ctgaacaact ataaacactg catcagcaac gttattgata acaagatcag cattgacgat
     4021 ctgggtagca tggataaagt tgacctgcac attctgaaca ccgcgtttca actgatcccg
     4081 gttgacaccg tgaacattga gcacaagaaa ctggttagcc tgatcgtgaa gcgtttcagc
     4141 accagcctgc tgagcagcgt tcgtgaggat cgtgtggact atgcgctgcg tcagagcttt
     4201 ctggaacgtt tcgcgtactt taccctgcat gcgccggtta gcgacatccc ggactacatt
     4261 aaaccgttcc tggatggttt taacggcagc gagccgatca gcgaactgtt caagaaattt
     4321 attctggtgg aggaccgtct gaacacctat gcgaagttct ggaaagtttg ggacctgttc
     4381 ttcgacaagg tggttaccct gtgcaaagat ggcgaccgtt actggtacgt tgacaagatc
     4441 attaaaagct acctgttcgc ggagagcccg tggaaggaaa acagcaacgg ctggcacacc
     4501 tttaaagaca gcaacagcca attcttttgc gatgttagcc gtaccatggg tcactgcccg
     4561 agcaccctgt atagcctggc gaagagcctg aacaacatcg cgagctgcta cctgaaccag
     4621 ggtatcacct ggctgagcga gattctgagc gtgaacaaga aactgtggga gaagaaactg
     4681 gaaaacgata ccgtttacta tctggaatgc ctggtgcgtc gttacatcaa caacgagcgt
     4741 gaacgtattc gtcgtaccaa gcagctgaaa caagaggttc tggtgatcct ggacttcctg
     4801 gttgaaaaag gtagcgtggt tggctatatg agccgtgaga acattctggg cggaagccat
     4861 caccatcatc accacagcag cggctaatta acctaggctg ctgccaccgc tgagcaataa
     4921 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag
     4981 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     5041 tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc atcgtggccg
     5101 gatcttgcgg cccctcggct tgaacgaatt gttagacatt atttgccgac taccttggtg
     5161 atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga
     5221 tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc
     5281 ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact
     5341 gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg
     5401 ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga
     5461 accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct
     5521 tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga
     5581 atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga
     5641 atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca
     5701 ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc
     5761 cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact
     5821 gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca
     5881 actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat actcttcctt
     5941 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6001 tgtatttaga aaaataaaca aatagctagc tcactcggtc gctacgctcc gggcgtgaga
     6061 ctgcggcggg cgctgcggac acatacaaag ttacccacag attccgtgga taagcagggg
     6121 actaacatgt gaggcaaaac agcagggccg cgccggtggc gtttttccat aggctccgcc
     6181 ctcctgccag agttcacata aacagacgct tttccggtgc atctgtggga gccgtgaggc
     6241 tcaaccatga atctgacagt acgggcgaaa cccgacagga cttaaagatc cccaccgttt
     6301 ccggcgggtc gctccctctt gcgctctcct gttccgaccc tgccgtttac cggatacctg
     6361 ttccgccttt ctcccttacg ggaagtgtgg cgctttctca tagctcacac actggtatct
     6421 cggctcggtg taggtcgttc gctccaagct gggctgtaag caagaactcc ccgttcagcc
     6481 cgactgctgc gccttatccg gtaactgttc acttgagtcc aacccggaaa agcacggtaa
     6541 aacgccactg gcagcagcca ttggtaactg ggagttcgca gaggatttgt ttagctaaac
     6601 acgcggttgc tcttgaagtg tgcgccaaag tccggctaca ctggaaggac agatttggtt
     6661 gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt ccccaactga cttaaccttc
     6721 gatcaaacca cctccccagg tggttttttc gtttacaggg caaaagatta cgcgcagaaa
     6781 aaaaggatct caagaagatc ctttgatctt ttctactgaa ccgctctaga tttcagtgca
     6841 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     6901 atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     6961 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     7021 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7081 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     7141 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     7201 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     7261 tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc
     7321 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     7381 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     7441 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     7501 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     7561 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     7621 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     7681 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     7741 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     7801 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     7861 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     7921 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     7981 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     8041 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     8101 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     8161 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     8221 aat
//
