LOCUS       V015773                 7213 bp    DNA     circular SYN 12-JUN-2021
DEFINITION  Exported.
ACCESSION   V015773
VERSION     V015773
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7213)
  AUTHORS   Robertson WE, Funke LFH., de la Torre D, Fredens J, Elliott TS,
            Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen
            S, Salmond GPC., Chin JW.
  TITLE     Sense Codon Reassignment Enables Viral Resistance and Encoded
            Polymer Synthesis
  JOURNAL   Science (2021) In press
REFERENCE   2  (bases 1 to 7213)
  AUTHORS   Robertson WE, Funke LFH., de la Torre D, Fredens J, Elliott TS,
            Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen
            S, Salmond GPC., Chin JW.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-APR-2021) Protein and Nucleic Acid Chemistry, MRC
            Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge,
            Cambridgeshire CB2 0QH, United Kingdom
REFERENCE   3  (bases 1 to 7213)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Science
            (2021) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (06-APR-2021) Protein and Nucleic Acid Chemistry, MRC Laboratory of
            Molecular Biology, Francis Crick Avenue, Cambridge, Cambridgeshire
            CB2 0QH, United Kingdom"
FEATURES             Location/Qualifiers
     source          1..7213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            41..65
                     /gene="glnS"
                     /label="glnS"
     regulatory      41..65
                     /gene="glnS"
                     /note="GlnS' promoter; GlnS' p"
                     /regulatory_class="promoter"
     CDS             110..1471
                     /gene="pylS"
                     /label="Pyrrolysine--tRNA ligase"
                     /note="Pyrrolysine--tRNA ligase from Methanosarcina mazei
                     (strain ATCC BAA-159 / DSM 3647 / Goe1 / Go1 / JCM 11833 /
                     OCM 88). Accession#: Q8PWY1"
     CDS             1535..1558
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     regulatory      2084..2134
                     /label="lpp promoter"
                     /note="lpp promoter"
                     /regulatory_class="promoter"
     tRNA            2141..2212
                     /product="tRNA-Ser"
                     /label="pylToptserU(CGA)"
                     /note="pylToptserU(CGA)"
     regulatory      2219..2247
                     /note="rrnC terminator; rrnC term"
                     /regulatory_class="terminator"
     terminator      2394..2480
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(2821..3366)
                     /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."
     promoter        3478..3506
                     /label="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        3554..3645
                     /label="AmpR promoter"
     CDS             3646..4446
                     /label="ApmR"
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
     CDS             complement(4760..5635)
                     /label="araC"
                     /note="L-arabinose regulatory protein"
     promoter        5662..5946
                     /label="araBAD promoter"
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite
                     direction (Guzman et al., 1995)"
     CDS             5986..6003
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             6004..6294
                     /label="SUMO"
                     /note="cleavable ubiquitin-like protein tag"
     CDS             6322..6915
                     /label="Mxe GyrA intein"
                     /note="modified GyrA intein (Southworth et al., 1999)"
     misc_feature    6937..6939
                     /label="TCA to AGT"
                     /note="TCA to AGT"
     CDS             6946..7101
                     /label="CBD"
                     /note="chitin binding domain from chitinase A1 (Watanabe et
                     al., 1994)"
     CDS             7112..7141
                     /label="Myc"
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             7157..7174
                     /label="6xHis"
                     /note="6xHis affinity tag"
ORIGIN
        1 gagagaagat tttcagcctg atacagatta aatcagaacg ttgtcagcct gtcccgcttt
       61 aatatcatac gccgttatac gttgtttacg ctttgaggaa tcccatatga tggacaaaaa
      121 accgctgaac accctgatct ctgctaccgg tctgtggatg tctcgtaccg gtaccatcca
      181 caaaatcaaa caccacgaag tttctcgttc taaaatctac atcgaaatgg cttgcggtga
      241 ccacctggtt gttaacaact ctcgttcttc tcgtaccgct cgtgctctgc gtcaccacaa
      301 ataccgtaaa acctgcaaac gttgccgtgt ttctgacgaa gacctgaaca aattcctgac
      361 caaagctaac gaagaccaga cctctgttaa agttaaagtt gtttctgctc cgacccgtac
      421 caaaaaagct atgccgaaat ctgttgctcg tgctccgaaa ccgctggaaa acaccgaagc
      481 tgctcaggct cagccgtctg gttctaaatt ctctccggct atcccggttt ctacccagga
      541 atctgtttct gttccggctt ctgtttctac ctctatctct tctatctcta ccggtgctac
      601 cgcttctgct ctggttaaag gtaacaccaa cccgatcacc tctatgtctg ctccggttca
      661 ggcttctgct ccggctctga ccaaatctca gaccgaccgt ctggaagttc tgctgaaccc
      721 gaaagacgaa atctctctga actctggtaa accgttccgt gaactggaat ctgaactgct
      781 gtctcgtcgt aaaaaagact tacaacagat ctacgctgaa gaacgtgaaa actacctggg
      841 taaactggaa cgtgaaatca cgcgtttctt cgttgaccgt ggtttcctgg aaatcaaatc
      901 tccgatcctg atcccgctgg aatacatcga acgtatgggt atcgacaacg acaccgaact
      961 gtctaaacag atcttccgtg ttgacaaaaa cttctgcctg cgtccgatgc tggctccgaa
     1021 cctgtacaac tacctgcgta aactggaccg tgctctgccg gacccgatca aaatcttcga
     1081 aatcggtccg tgctaccgta aagaatctga cggtaaagaa cacctggaag aattcaccat
     1141 gctgaacttc tgccagatgg gttctggttg cacccgtgaa aacctggaat ctatcatcac
     1201 cgacttcctg aaccacctgg gtatcgactt caaaatcgtt ggtgactctt gcatggttta
     1261 cggtgacacc ctggacgtta tgcacggtga cctggaactg tcttctgctg ttgttggtcc
     1321 gatcccgctg gaccgtgaat ggggtatcga caaaccgtgg atcggtgctg gtttcggtct
     1381 ggaacgtctg ctgaaagtta aacacgactt caaaaacatc aaacgtgctg ctcgttctga
     1441 atcttactac aacggtatct ctaccaacct gtctggtggc ggtggctctg gtggcggtgg
     1501 ctctggtggc ggtggctctg gtggcggtgg ctctgactac aaggacgacg acgacaagta
     1561 actgcagttt caaacgctaa attgcctgat gcgctacgct tatcaggcct acatgatctc
     1621 tgcaatatat tgagtttgcg tgcttttgta ggccggataa ggcgttcacg ccgcatccgg
     1681 caagaaacag caaacaatcc aaaacgccgc gttcggcggt cgacacagat gtaggtgttc
     1741 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctt
     1801 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gctgccggca
     1861 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
     1921 cccgcgccca ccggaaggag ctaccggcag cggtgcggac tgttgtaact cagaataaga
     1981 aatgaggccg ctcatggcgt tctgttgccc gtctcactgg tgaaaagaaa aacaaccctg
     2041 gcgccgcttc tttgagcgaa cgatcaaaaa taagtggcgc cccatcaaaa aaatattctc
     2101 aacataaaaa actttgtgta atacttgtaa cgctagatct ggaaacgtga tcatgtagat
     2161 cgaacggtct cgaaaaccgt tcagtggggt tagattcccc acgtttccgc caactagtat
     2221 ccttagcgaa agctaaggat tttttttcag aagcggtctg ataaaacaga atttgcctgg
     2281 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     2341 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     2401 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     2461 ctctcctgag taggacaaat ccgccgggag ctgtccctcc tgttcagcta ctgacggggt
     2521 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
     2581 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     2641 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     2701 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     2761 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     2821 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     2881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     2941 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     3001 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     3061 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3121 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     3181 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     3241 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     3301 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     3361 tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc
     3421 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgtttg
     3481 acagcttatc atcgataagc tttaatgcgg tagtttatca cagttaaatt gctaacgcag
     3541 tcaggcaccg tgttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3601 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tagtgcggtg
     3661 gagtgcaatg tcgtgcaata cgaatggcga aaagccgagc tcatcggtca gcttctcaac
     3721 cttggggtta cccccggcgg tgtgctgctg gtccacagct ccttccgtag cgtccggccc
     3781 ctcgaagatg ggccacttgg actgatcgag gccctgcgtg ctgcgctggg tccgggaggg
     3841 acgctcgtca tgcccagctg gagtggtctg gacgacgagc cgttcgatcc tgccacgagc
     3901 cccgttacac cggaccttgg agttgtctct gacacattct ggcgcctgcc aaatgtaaag
     3961 cgcagcgccc atccatttgc ctttgcggca gcggggccac aggcagagca gatcatctct
     4021 gatccattgc ccctgccacc tcacagccct gcaagcccgg tcgcccgtgt ccatgaactc
     4081 gatgggcagg tacttctcct cggcgtggga cacgatgcca acacgacgct gcatcttgcc
     4141 gagttgatgg caaaggttcc ctatggggtg ccgagacact gcaccattct tcaggatggc
     4201 aagttggtac gcgtcgatta tctcgagaat gaccactgct gtgagcgctt tgccttggcg
     4261 gacaggtggc tcaaggagaa gagccttcag aaggaaggtc cagtcggtca tgcctttgct
     4321 cggttgatcc gctcccgcga cattgtggcg acagccctgg gtcaactggg ccgagatccg
     4381 ttgatcttcc tgcatccgcc agaggcggga tgcgaagaat gcgatgccgc tcgccagagc
     4441 attggctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca agaattggag
     4501 ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag aacatatcca
     4561 tcgcgtgcgg ccgcgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4621 aacgcgcgag gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct
     4681 gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg
     4741 tctgattcgt taccaattaa ctcaacttga cggctacatc attcactttt tcttcacaac
     4801 cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgg ctgaaataga
     4861 gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca
     4921 aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta
     4981 actgctggcg gaaaagatga ctcagacggc tcgggctcaa gcaaacatgc tgtgcgacgc
     5041 tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta
     5101 cccgattatc catcggtgga tggaggctct cgttaatcgc ttccatgcgc cgcagtaaca
     5161 attgctcaag cagatttatc gccagcagct cgctatagcg cccttcccct tgcccggcgt
     5221 taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga
     5281 accccgtatt ggcaaatata ctcggccagt taagccattc atgccagtag gcgcgcggac
     5341 gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa
     5401 tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat
     5461 ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg
     5521 gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca
     5581 gatgggcatt aaagctgtat cccggcagca ggggatcatt ttgcgcttca gccatacttt
     5641 tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc
     5701 actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca
     5761 ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc
     5821 acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat
     5881 ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt
     5941 ctccataccc gttttttggg ctaacaggag gaattaacca tggcacatca tcaccatcac
     6001 cacagcgaca gcgaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct
     6061 gagactcaca tcaatttaaa ggtgtccgat ggatctagtg agatcttctt caagatcaaa
     6121 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg
     6181 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat
     6241 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggttgcggt
     6301 tctgggtctg gttctgggtc ttgcatcacg ggagatgcac tagttgccct acccgagggc
     6361 gagagcgtac gcatcgccga catcgtgccg ggtgcgcggc ccaacagtga caacgccatc
     6421 gacctgaaag tccttgaccg gcatggcaat cccgtgctcg ccgaccggct gttccactcc
     6481 ggcgagcatc cggtgtacac ggtgcgtacg gtcgaaggtc tgcgtgtgac gggcaccgcg
     6541 aaccacccgt tgttgtgttt ggtcgacgtc gccggggtgc cgaccctgct gtggaagctg
     6601 atcgacgaaa tcaagccggg cgattacgcg gtgattcaac gcagcgcatt cagcgtcgac
     6661 tgtgcaggtt ttgcccgcgg gaaacccgaa tttgcgccca caacctacac agtcggcgtc
     6721 cctggactgg tgcgtttctt ggaagcacac caccgagacc cggacgccca agctatcgcc
     6781 gacgagctga ccgacgggcg gttctactac gcgaaagtcg ccagtgtcac cgacgccggc
     6841 gtgcagccgg tgtatagcct tcgtgtcgac acggcagacc acgcgtttat cacgaacggg
     6901 ttcgtcagcc acgctactgg cctcaccggt ctgaacagtg gcctcacgac aaatcctggt
     6961 gtatccgctt ggcaggtcaa cacagcttat actgcgggac aattggtcac atataacggc
     7021 aagacgtata aatgtttgca gccccacacc tccttggcag gatgggaacc atccaacgtt
     7081 cctgccttgt ggcagcttca atgatgggcc cgaacaaaaa ctcatctcag aagaggatct
     7141 gaatagcgcc gtcgaccatc atcatcatca tcattgagtt taaacggtct ccagcttggc
     7201 tgttttggcg gat
//
