LOCUS       V015699                 5524 bp    DNA     circular SYN 12-JUN-2021
DEFINITION  Exported.
ACCESSION   V015699
VERSION     V015699
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5524)
  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 5524)
  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 5524)
  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..5524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..48
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(222..1034)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(1035..1126)
                     /label="AmpR promoter"
     rep_origin      complement(1190..1779)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     gene            1863..1888
                     /gene="glnS"
                     /label="glnS"
     regulatory      1863..1888
                     /gene="glnS"
                     /label="GlnS promoter"
                     /note="GlnS promoter"
                     /regulatory_class="promoter"
     CDS             1933..3294
                     /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             3358..3381
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3434..4261
                     /codon_start=1
                     /transl_table=11
                     /product="PylRS variant 1R26 (Y126G-M129L)"
                     /label="PylRS variant 1R26"
                     /note="1R26MaPylRS; specific for CbzK; from
                     Methanomethylophilus alvus"
                     /protein_id="QWF36673.1"
                     /translation="MAEHFTDAQIQRLREYGNGTYKDMEFADVSAREKAFTKLMSDASR
                     DNESALKGMIAHPARQGLSRLMNDIADALVADGFIEVRTPIIISKDALAKMTITPDKPL
                     FKQVFWIDDKRALRPMLAPSLGTVLRSLRDHTDGPVKIFEMGSCFRKESHSGMHLEEFT
                     MLNLVDMGPAGDATESLKKYIGIVMKAAGLPDYQLVHEESDVYKETIDVEINGQEVCSA
                     AVGPHYLDAAHDVHEPWAGAGFGLERLLTIRQGYSTVMKGGASTTYLNGAKMD"
     misc_feature    3578..3580
                     /label="TCA to AGT"
                     /note="TCA to AGT"
     misc_feature    3623..3625
                     /label="TCA to AGT"
                     /note="TCA to AGT"
     misc_feature    3809..3811
                     /label="Y126G"
                     /note="Y126G"
     misc_feature    3818..3820
                     /label="M129L"
                     /note="M129L"
     misc_feature    3890..3892
                     /label="TCG to AGC"
                     /note="TCG to AGC"
     misc_feature    4094..4096
                     /label="TCG to AGC"
                     /note="TCG to AGC"
     regulatory      4784..4834
                     /label="lpp promoter"
                     /note="lpp promoter"
                     /regulatory_class="promoter"
     tRNA            4841..4912
                     /product="tRNA-Ser"
                     /label="pylToptserU(CGA)"
                     /note="pylToptserU(CGA)"
     tRNA            4952..5023
                     /product="tRNA-Pyl"
                     /label="Ma tRNA Pyl(8)(CUA)"
                     /note="Ma tRNA Pyl(8)(CUA)"
     gene            5030..5058
                     /gene="rrnC"
                     /label="rrnC"
     regulatory      5030..5058
                     /gene="rrnC"
                     /label="rrnC term"
                     /note="rrnC term"
                     /regulatory_class="terminator"
     primer_bind     complement(5068..5084)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             5429..5473
                     /label="S-Tag"
                     /note="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
ORIGIN
        1 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag
       61 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
      121 tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc atcgtggccg
      181 gatcttgcgg cccctcggct tgaacgaatt gttagacatt agaaaaactc atcgagcatc
      241 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
      301 ttctgtaaac taggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
      361 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
      421 ataaggttat caagactgaa atcaccatga gtgacgacac tatccggact gaatggcaaa
      481 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
      541 tcactcgcat caaccaaacc gttattcatt cgactttgcg cctgagcgag acgaaatacg
      601 cggtcgctgt taaaaggaca attacaaaca ggaatgctat gcaaccggcg caggaacact
      661 gccagcgcat caacaatatt ttcaccacta tcaggatatt cttctaatac ctggaatgct
      721 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
      781 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
      841 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
      901 ccatacaatc gatagattgt cgcaccactt tgcccgacat tatcgcgagc ccatttatac
      961 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     1021 tgaatatggc tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1081 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagc tagcgagcaa
     1141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1621 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt tttttttgtt
     1741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggg gatcctcggg
     1861 agttgtcagc ctgtcccgct tataagatca tacgccgtta tacgttgttt acgctttgag
     1921 gaatcccata tgatggacaa aaaaccgctg aacaccctga tctctgctac cggtctgtgg
     1981 atgtctcgta ccggtaccat ccacaaaatc aaacaccacg aagtttctcg ttctaaaatc
     2041 tacatcgaaa tggcttgcgg tgaccacctg gttgttaaca actctcgttc ttctcgtacc
     2101 gctcgtgctc tgcgtcacca caaataccgt aaaacctgca aacgttgccg tgtttctgac
     2161 gaagacctga acaaattcct gaccaaagct aacgaagacc agacctctgt taaagttaaa
     2221 gttgtttctg ctccgacccg taccaaaaaa gctatgccga aatctgttgc tcgtgctccg
     2281 aaaccgctgg aaaacaccga agctgctcag gctcagccgt ctggttctaa attctctccg
     2341 gctatcccgg tttctaccca ggaatctgtt tctgttccgg cttctgtttc tacctctatc
     2401 tcttctatct ctaccggtgc taccgcttct gctctggtta aaggtaacac caacccgatc
     2461 acctctatgt ctgctccggt tcaggcttct gctccggctc tgaccaaatc tcagaccgac
     2521 cgtctggaag ttctgctgaa cccgaaagac gaaatctctc tgaactctgg taaaccgttc
     2581 cgtgaactgg aatctgaact gctgtctcgt cgtaaaaaag acttacaaca gatctacgct
     2641 gaagaacgtg aaaactacct gggtaaactg gaacgtgaaa tcacgcgttt cttcgttgac
     2701 cgtggtttcc tggaaatcaa atctccgatc ctgatcccgc tggaatacat cgaacgtatg
     2761 ggtatcgaca acgacaccga actgtctaaa cagatcttcc gtgttgacaa aaacttctgc
     2821 ctgcgtccga tgctggctcc gaacctgtac aactacctgc gtaaactgga ccgtgctctg
     2881 ccggacccga tcaaaatctt cgaaatcggt ccgtgctacc gtaaagaatc tgacggtaaa
     2941 gaacacctgg aagaattcac catgctgaac ttctgccaga tgggttctgg ttgcacccgt
     3001 gaaaacctgg aatctatcat caccgacttc ctgaaccacc tgggtatcga cttcaaaatc
     3061 gttggtgact cttgcatggt ttacggtgac accctggacg ttatgcacgg tgacctggaa
     3121 ctgtcttctg ctgttgttgg tccgatcccg ctggaccgtg aatggggtat cgacaaaccg
     3181 tggatcggtg ctggtttcgg tctggaacgt ctgctgaaag ttaaacacga cttcaaaaac
     3241 atcaaacgtg ctgctcgttc tgaatcttac tacaacggta tctctaccaa cctgtctggt
     3301 ggcggtggct ctggtggcgg tggctctggt ggcggtggct ctggtggcgg tggctctgac
     3361 tacaaggacg acgacgacaa gtaacctatt gaagggttgc taagaataac aagatcaaat
     3421 aaggaggttt tttatggcag aacattttac agatgcacag attcagcgct tacgcgagta
     3481 tggtaatggg acatataagg atatggagtt cgcagacgtg agtgcgcgtg aaaaggcgtt
     3541 cacgaagctt atgagtgatg ccagtcgcga taatgaaagt gcccttaagg gaatgattgc
     3601 gcaccctgca cgccagggtt taagtcgctt aatgaatgac attgcagacg ctttggtggc
     3661 tgatggattt attgaagttc gcacacccat tattatcagt aaagatgctt tagccaaaat
     3721 gacaatcact cccgataaac cattattcaa acaagtattt tggattgatg ataaacgcgc
     3781 tctgcgtcca atgttagccc cgagtctggg cacggtcctg cgtagcctgc gtgatcatac
     3841 tgacggccct gtcaaaattt tcgaaatggg cagttgtttt cgcaaagaaa gccacagtgg
     3901 gatgcacctg gaagagttta ccatgttaaa cctggtagat atgggccccg ccggcgatgc
     3961 cactgagagc ctgaagaaat atattgggat cgtaatgaag gccgccgggc tgcctgacta
     4021 ccaattagtc cacgaggaaa gtgatgtgta taaagaaacg attgatgttg aaatcaacgg
     4081 acaagaggtt tgtagcgctg ctgtaggtcc tcactatttg gatgccgccc atgacgtgca
     4141 cgagccgtgg gcaggggcag ggttcggcct ggagcgcctg ctgacaattc gccagggata
     4201 tagcacagtg atgaaagggg gagctagcac aacctatttg aacggggcta agatggacta
     4261 actgcagttt caaacgctaa attgcctgat gcgctacgct tatcaggcct acatgatctc
     4321 tgcaatatat tgagtttgcg tgcttttgta ggccggataa ggcgttcacg ccgcatccgg
     4381 caagaaacag caaacaatcc aaaacgccgc gttcggcggt cgacacagat gtaggtgttc
     4441 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctt
     4501 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gctgccggca
     4561 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
     4621 cccgcgccaa ccggaaggag ctaccggcag cggtgcggac tgttgtaact cagaataaga
     4681 aatgaggccg ctcatggcgt tctgttgccc gtctcactgg tgaaaagaaa aacaaccctg
     4741 gcgccgcttc tttgagcgaa cgatcaaaaa taagtggcgc cccatcaaaa aaatattctc
     4801 aacataaaaa actttgtgta atacttgtaa cgctagatct ggaaacgtga tcatgtagat
     4861 cgaacggtct cgaaaaccgt tcagtggggt tagattcccc acgtttccgc caaattttgc
     4921 acccagcaaa cttggtacgt aaacgcatcg tgggggacgg tccggcgacc agcgggtctc
     4981 taaaacctag ccttgcgggg ttcgacaccc cggtctctcg ccaactagta tccttagcga
     5041 aagctaagga ttttttttaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga
     5101 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagacgctc
     5161 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     5221 cgccgccgca aggaatggtg catgcaagga gcccgagatg cgccgcgtgc ggctgctgga
     5281 gatggcggac gcgatggata tgttctggcg gccgcataat gcttaagtcg aacagaaagt
     5341 aatcgtattg tacacggccg cataatcgaa attaaattgg atatcggcag gccacgcgat
     5401 cgctgacgtc ggtaccctcg agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca
     5461 gcacatggac tcgtctacta gcgcagctta attaacctag gctgctgcca ccgctgagca
     5521 ataa
//
