LOCUS       sequence-369977-        8733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon optimized CcAvs4 with C-terminal His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8733)
  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 8733)
  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-CcAvs4
FEATURES             Location/Qualifiers
     source          1..8733
                     /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             5368..5385
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5417..5435)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5431..5478
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5649..6440)
                     /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(6441..6532)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6580..7318)
                     /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(7528..8610)
                     /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     8572..8591
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8611..8688)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg caccaggcag ctcaagtatc aaggcccgtt
      121 gattttgtta ttattgcggc tttggaacag gagcgggacg caatgttgcg acatctgccg
      181 ggctacagcc ggctggaccg tgaggacgac gatccgcata cctattatga agcgttcgtg
      241 ccgaccatgc gttccgatgg tgcgcgatac cgtgttcttg ttaccagctt agctggcatg
      301 ggtccgattc aagctgcggc gaaggctatg gcggtcgttc accgctgggg tccgagacac
      361 gtgctgttcg tgggcatcgc cggcggtcgt cggtcggtgg caggccttgg tgacgtgctg
      421 gttgctaacc aaatagcgga ctacaccgtg ggtaagcgtc ttgaagatag gcgtattacc
      481 cgttgggagg tgtaccgtgt ggacgccact ctcttggacg cagccatcaa cctgccgggt
      541 ggttgggaaa gccgcaccca ggaggtgcgt ccggttcccg gtcgcccgtt gaggcatagt
      601 ggcattatcc tgtcgggtgg tgacgtgctg gccgatggtg agaccctgcg tcagaatcag
      661 gaagtatggg ataaactgat cggcgtcgag atggaaggcg gaggcaccgc gctagccctg
      721 caccagaccg gctcccgtcc gcggttccta atgattcgtg gtgttagcga tcttgcggac
      781 gctgagaagg gtagcgctac cgtcgagaac tggcgtccat acgcctgcga cgtcgccgcg
      841 gcgtacaccc gcgcgttctt ggagtccggc ccggtcccgg ctagttcggg cccgagcgac
      901 ggcgctggta gcggagattg gaccactttg gccaacctga ccacagatta taaagcacac
      961 atcgaggcgt ccctgccgaa cggtgtttct attgcgagac acgatgaagc tggtaagctg
     1021 gaggccgcgc tggccgagaa ctctatcagt gtcgttttcg gtgactctgg tgcgggtaag
     1081 tcggctctgg tcaaaaccgt tttggacacg cgttttgcag cgtggaccca ggtttggttg
     1141 ggtccgaacc aggttcaggc agccctgagc gtggtgcgtc gttccagcct gccgctgcag
     1201 catgcactgc tggatgtgct caaagccagc gctaacgcac gcaacgtgct ggtcttggat
     1261 agcgtagagc gtatcgaacc gtccgaaacg agcattgttc gcaggtttgt tcatgaattg
     1321 ctagcaagcg ataatagccc gcaggctgct tggcgcctga tcgtgattag ccaaacccag
     1381 agctgggcgg aacgtgctga gacaatcctg ggcgcccaac cgagcgttct gatcgaacta
     1441 ggcctgttga aaagttctga tgtcgaggcg gcgctgtccg ccaccccagc gctgtcttgg
     1501 ttagttggtc acccggaaac cgttgcggtt ctggccaacc ttaagacgct tggctgggtt
     1561 attcaggcaa gggctgctct gacagcgggc accgtggccc ctgcctccca cgcagcggtt
     1621 gccgaccgac tgtggcgttt ctggactgac ggccgcgcgg agattcaaga gctgcttatg
     1681 cgcctggccg aacgtgaggc aagctttgaa cgctcgtttg cactgacgga cctgcgtccg
     1741 ggggatgcaa gaacctttca gcagggtcct gcgcacctgc cgctccgctt gaatgaacgt
     1801 accaatcacc tggagtttga acatgatctg gcggctgatt gggcgcgctt ccagtttcta
     1861 aagcaaatcg cgcgtaacac cgctcgctgg tcggcgctag ccagcaatcc gctgtggtca
     1921 gctgctttac gtatgcttgg ccagttcctg ctgcgtcaac cggcgggcac cgcgactgct
     1981 tgggatgaag cattcgccgc ggcggaggaa actgaaatgc cgctggctag cgacatgctg
     2041 ttagatgccc tttgcctgga tccggaagcg gaacgcttcc tgtacgaacg tgttgacttg
     2101 ttgttagcag agaacggtaa gcgtatgacc cgtcttttga ccagattcca tcatataggt
     2161 accgtcccgc gtgttctccc accgctgagc gacaacccga ccctaggcct tcacattgaa
     2221 actcgttacc gcagtgtaat agcggctcgt tggcctccaa ttgtacgttt cttggcgacc
     2281 catagggatc gcattacgtc tatgctactg ccggcgttag caaagctggc ggagacctgg
     2341 ctgtccggca ccccgccgat gctagcgaat ggcaacccga ccccgttccg ccgtgagttc
     2401 gctgaattgg cgattgcaat ggcacgcacc gtgcaggtgc tcaaggaaca aggtgttatg
     2461 tttctctccc aagatccggt gttttttgcg gctccactcg cgggagcgcc agatctgcct
     2521 gacgaagtca gctgttgggc gttagagatg agcggcagac ggccgatctc aaaagatgtc
     2581 ttgatcagag tggcggtggc ccgcaaacgt caagcggagg aacgtgctgc gcgtttgaag
     2641 aacgacaccc gtatgcgtgc gaccgaatca cgtcgtcgtg acctgccgca gttggtggta
     2701 ccggggcgcc acgaactgga accgtggcct ctgggtgcca tgcgtccgat cgatcatgat
     2761 tttcgtaacg cgtgcctggg tggcagctcg ctaggcccat taatgcgtac ccgtccgacg
     2821 gttgcggcag aggttctgct ggcgctgatc atcaaggatc agccgtgtga agaatatggt
     2881 gcaggtcgtt ggaaaatcaa attaggcctg gaagcggcac gtgatggcta cccgaccgcg
     2941 ttctggaaaa ccccgttctt cctgttcctg caaattgccc cgcgtgaagc gcttcaagcg
     3001 atcatcaaac tggtgaattt ctgcaccgat cgctgggtgg atagtgcacg gttcggcacc
     3061 aaccgttctg tgccaacggt gaaattggag atgggtaacg gtattgagaa agagtttatc
     3121 ggaggcccac tgctgtttgg ttggacgcag gaaaatagca tccgcgacgg caaccttcac
     3181 tgtgcacttg acgcgctgga gcgctggctg acattgcagg ttgacaaaaa cgccgacatc
     3241 gcgcattggg ttgagggtat tttgaaagaa ggagcctccg tggcgttctt gggcctcctc
     3301 gttaatctgg gtaagtatca gccggcgctg ttgccgggtc cgcttgctcc gctgctgacc
     3361 agccccctga tgttttactg ggatgcccag tgcgttgaac aagttggcag ccgtttcatc
     3421 ggttggcatt gggctaatag cagcgaagtg atgttcgagt tcgccaaatc atggaccttg
     3481 gcggcacaca gacgtcagag attcgtggat atcgtggcga atctgctgaa ggaagacgac
     3541 gtcgttgcgg aaaagctgca gtttgcgatc ccgacatggg taatcccgaa cgacccgaaa
     3601 cagtctctgg agctgcgttt catgtgggca gcgttagaca gggcgaatta tcatcgtgtc
     3661 atcgacagcg aaaccggcgc cgaagtcttc gccttggtgt acccggaaag cctgcgtcag
     3721 gaggcaaagg catggcagaa aagccacgaa gccgagatcc aacaattgat tatcccgagc
     3781 cagtgcgaac agctgctgcg tgcgcagcaa agcttgagtg atgacgttgc gggccgtttc
     3841 tatgagctgc tagagtcctc taaccgtagc agcaccgtag aatcttccgt aaagatgaaa
     3901 agcatgctcg ctctggccac gaccttggtg gtttgcggcg cgtcctggct gggtcgtgaa
     3961 ccaacgaaac gccaaagact ggtggaaatc atccgcgcta gcgcgctcga ggtgggcgag
     4021 accgcggttg ctatccgtac ggcccgcatc tccgggggtg aggactccct gccgtttacc
     4081 gcctttgcgg tgatgcacct gtggaccaaa tttgatgaga gcgcggagtg ggaagcgctc
     4141 gtcctccgct tactgacgtg cggtgactcg cgtgttgtcg ttacactggt ttccatcgca
     4201 tatgctaagc gtgagatgct aggcagcacc tggtggcgct tcctgcaagt tggtctgttt
     4261 tggagcgcgc tggttatgct ggcacccaac tatgacgatt ccgagcgtat cggtccgatt
     4321 tggcacggct ggctggcgcg cattcgtcgc ttcccgctac gtcaagaggg tgcggtttct
     4381 acgggcgtta agatcggccg cgttttaatg ggctaccagc gtctggagcg agcacgttgg
     4441 gcgagagcga acggtctgac cgtccagcaa tgccgtggtc gtgagttcga cgctcgccac
     4501 agcatcggtt tagacattaa ctttctgggt ctccttttca actggctgtt gaagggtctg
     4561 ggcacgggcg acctgctgaa ggaccgcgaa ctggtgctgc agctatggga ttgtgaagtt
     4621 gagtacatgc aggcccaggg tgaagctgac ggtgaatgca gcccgccgtc ccaacaaata
     4681 ggttacgatt tgctggcgaa gttgggctcc ttaagcgtgt cactgagtcc ggcggaggcg
     4741 cgtccggtct gggaggcggt gttatcttgc ggtccgaaag cgcactatgc ccttcaatat
     4801 tttacccgca acctgttttt actcctgaat caaggttgtg atgttgagcg attccgtcct
     4861 ctgtggcgtt ctttggttca gtatgcactg gacgctaagt ggggtgcacg tgatctgtac
     4921 tggccgtacg gcgagcgttt gctcggcgac ctcctgggct ttggcaacga agatctgctg
     4981 aagcacctgc cttcaaaatc cgtgccggat atgcgtgacc tgtatgatcg atgggcacaa
     5041 gcacacctgg gtcatagcga ggagtccgtg cggcgtttct gccgtttcct gagctccgag
     5101 gtggggactt ttctgcgtct cgacggtctg cgttggatta gcgcggcgct acgtgtgagc
     5161 aaaaccggta gacaatggtg gcgcgatgaa aatggcgacg ccttggttga actgctggac
     5221 accgcactga cgcgtaatgc cccggcattg gcgaacgacg cgaaggcgcg tgaagctctg
     5281 gtggagctcg ctgctgagct ggctgctcgt aacattccga ttgccttgac cttacaggag
     5341 agaattcgtc tgtttcgtgg cggaagccat caccatcatc accacagcag cggctaatta
     5401 acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     5461 gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt
     5521 ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc
     5581 cctgaaccga cgaccgggtc atcgtggccg gatcttgcgg cccctcggct tgaacgaatt
     5641 gttagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct
     5701 tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta
     5761 gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg
     5821 acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc
     5881 actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca
     5941 tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga
     6001 cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg
     6061 atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc
     6121 agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact
     6181 tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg
     6241 atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata
     6301 tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac
     6361 gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg
     6421 gcgatcaccg cttccctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6481 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aatagctagc
     6541 tcactcggtc gctacgctcc gggcgtgaga ctgcggcggg cgctgcggac acatacaaag
     6601 ttacccacag attccgtgga taagcagggg actaacatgt gaggcaaaac agcagggccg
     6661 cgccggtggc gtttttccat aggctccgcc ctcctgccag agttcacata aacagacgct
     6721 tttccggtgc atctgtggga gccgtgaggc tcaaccatga atctgacagt acgggcgaaa
     6781 cccgacagga cttaaagatc cccaccgttt ccggcgggtc gctccctctt gcgctctcct
     6841 gttccgaccc tgccgtttac cggatacctg ttccgccttt ctcccttacg ggaagtgtgg
     6901 cgctttctca tagctcacac actggtatct cggctcggtg taggtcgttc gctccaagct
     6961 gggctgtaag caagaactcc ccgttcagcc cgactgctgc gccttatccg gtaactgttc
     7021 acttgagtcc aacccggaaa agcacggtaa aacgccactg gcagcagcca ttggtaactg
     7081 ggagttcgca gaggatttgt ttagctaaac acgcggttgc tcttgaagtg tgcgccaaag
     7141 tccggctaca ctggaaggac agatttggtt gctgtgctct gcgaaagcca gttaccacgg
     7201 ttaagcagtt ccccaactga cttaaccttc gatcaaacca cctccccagg tggttttttc
     7261 gtttacaggg caaaagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7321 ttctactgaa ccgctctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
     7381 cagccccata cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag
     7441 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     7501 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     7561 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     7621 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     7681 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     7741 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     7801 taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     7861 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     7921 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     7981 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     8041 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     8101 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     8161 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     8221 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     8281 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     8341 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     8401 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     8461 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     8521 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     8581 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     8641 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     8701 gctctccctt atgcgactcc tgcattagga aat
//
