LOCUS       V015808                 5114 bp    DNA     circular SYN 12-JUN-2021
DEFINITION  Exported.
ACCESSION   V015808
VERSION     V015808
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5114)
  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 5114)
  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 5114)
  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..5114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             197..241
                     /label="S-Tag"
                     /note="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
     terminator      293..340
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(514..1326)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(1327..1418)
                     /label="AmpR promoter"
     rep_origin      complement(1482..2071)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     gene            2155..2180
                     /gene="glnS"
                     /label="glnS"
     regulatory      2155..2180
                     /gene="glnS"
                     /label="GlnS promoter"
                     /note="GlnS promoter"
                     /regulatory_class="promoter"
     CDS             2222..3049
                     /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="QWF36669.1"
                     /translation="MAEHFTDAQIQRLREYGNGTYKDMEFADVSAREKAFTKLMSDASR
                     DNESALKGMIAHPARQGLSRLMNDIADALVADGFIEVRTPIIISKDALAKMTITPDKPL
                     FKQVFWIDDKRALRPMLAPSLGTVLRSLRDHTDGPVKIFEMGSCFRKESHSGMHLEEFT
                     MLNLVDMGPAGDATESLKKYIGIVMKAAGLPDYQLVHEESDVYKETIDVEINGQEVCSA
                     AVGPHYLDAAHDVHEPWAGAGFGLERLLTIRQGYSTVMKGGASTTYLNGAKMD"
     misc_feature    2366..2368
                     /label="TCA to AGT"
                     /note="TCA to AGT"
     misc_feature    2411..2413
                     /label="TCA to AGT"
                     /note="TCA to AGT"
     misc_feature    2597..2599
                     /label="Y126G"
                     /note="Y126G"
     misc_feature    2606..2608
                     /label="M129L"
                     /note="M129L"
     misc_feature    2678..2680
                     /label="TCG to AGC"
                     /note="TCG to AGC"
     misc_feature    2882..2884
                     /label="TCG to AGC"
                     /note="TCG to AGC"
     CDS             3099..4067
                     /gene="tyrS"
                     /label="Tyrosine--tRNA ligase"
                     /note="Tyrosine--tRNA ligase from Archaeoglobus fulgidus
                     (strain ATCC 49558 / DSM 4304 / JCM 9628 / NBRC 100126 /
                     VC-16). Accession#: O29482"
     regulatory      4663..4713
                     /label="lpp promoter"
                     /note="lpp promoter"
                     /regulatory_class="promoter"
     tRNA            4720..4791
                     /product="tRNA-Pyl"
                     /label="Ma tRNA Pyl(8)(CGA)"
                     /note="Ma tRNA Pyl(8)(CGA)"
     tRNA            4831..4905
                     /product="tRNA-Tyr"
                     /label="Af tRNA TyrA01(CUA)"
                     /note="Af tRNA TyrA01(CUA)"
     gene            4912..4940
                     /gene="rrnC"
                     /label="rrnC"
     regulatory      4912..4940
                     /gene="rrnC"
                     /label="rrnC term"
                     /note="rrnC term"
                     /regulatory_class="terminator"
     primer_bind     complement(4950..4966)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
ORIGIN
        1 gaatggtgca tgcaaggagc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc
       61 gatggatatg ttctggcggc cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta
      121 cacggccgca taatcgaaat taaattggat atcggcaggc cacgcgatcg ctgacgtcgg
      181 taccctcgag tctggtaaag aaaccgctgc tgcgaaattt gaacgccagc acatggactc
      241 gtctactagc gcagcttaat taacctaggc tgctgccacc gctgagcaat aactagcata
      301 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaacctc aggcatttga
      361 gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata
      421 agcggctatt taacgaccct gccctgaacc gacgaccggg tcatcgtggc cggatcttgc
      481 ggcccctcgg cttgaacgaa ttgttagaca ttagaaaaac tcatcgagca tcaaatgaaa
      541 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
      601 actaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
      661 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
      721 atcaagactg aaatcaccat gagtgacgac actatccgga ctgaatggca aaagtttatg
      781 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
      841 atcaaccaaa ccgttattca ttcgactttg cgcctgagcg agacgaaata cgcggtcgct
      901 gttaaaagga caattacaaa caggaatgct atgcaaccgg cgcaggaaca ctgccagcgc
      961 atcaacaata ttttcaccac tatcaggata ttcttctaat acctggaatg ctgttttccc
     1021 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     1081 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     1141 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     1201 tcgatagatt gtcgcaccac tttgcccgac attatcgcga gcccatttat acccatataa
     1261 atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg
     1321 gctcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1381 cggatacata tttgaatgta tttagaaaaa taaacaaata gctagcgagc aaaaggccag
     1441 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1501 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1561 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1621 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1681 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1741 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1801 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1861 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1921 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1981 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gttttttttg tttgcaagca
     2041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2101 tgacgctcag tggaacgaaa actcacgtta agggattttg gggatcctcg ggagttgtca
     2161 gcctgtcccg cttataagat catacgccgt tatacgttgt ttacgctttg aggaatccca
     2221 tatggcagaa cattttacag atgcacagat tcagcgctta cgcgagtatg gtaatgggac
     2281 atataaggat atggagttcg cagacgtgag tgcgcgtgaa aaggcgttca cgaagcttat
     2341 gagtgatgcc agtcgcgata atgaaagtgc ccttaaggga atgattgcgc accctgcacg
     2401 ccagggttta agtcgcttaa tgaatgacat tgcagacgct ttggtggctg atggatttat
     2461 tgaagttcgc acacccatta ttatcagtaa agatgcttta gccaaaatga caatcactcc
     2521 cgataaacca ttattcaaac aagtattttg gattgatgat aaacgcgctc tgcgtccaat
     2581 gttagccccg tctctgggca cggtcctgcg tagcctgcgt gatcatactg acggccctgt
     2641 caaaattttc gaaatgggct cttgttttcg caaagaaagc cacagtggga tgcacctgga
     2701 agagtttacc atgttaaacc tggtagatat gggccccgcc ggcgatgcca ctgagtccct
     2761 gaagaaatat attgggatcg taatgaaggc cgccgggctg cctgactacc aattagtcca
     2821 cgaggaatct gatgtgtata aagaaacgat tgatgttgaa atcaacggac aagaggtttg
     2881 tagcgctgct gtaggtcctc actatttgga tgccgcccat gacgtgcacg agccgtgggc
     2941 aggggcaggg ttcggcctgg agcgcctgct gacaattcgc cagggatata gcacagtgat
     3001 gaaaggggga gcttccacaa cctatttgaa cggggctaag atggactaac ctattgaagg
     3061 gttgctaaga ataacaagat caaataagga ggttttttat ggacattacg gaaaaactgc
     3121 gcttgattac gcgcaacgcg gaggaagtcg taacagaaga agagttgcgt cagttgattg
     3181 aaacaaaaga aaaaccccgt gcaattgtag gctatgaacc aagtggcgag atccatttag
     3241 gtcatatgat gaccgtccag aaactgatgg atttacaaga ggcgggattt gagattattg
     3301 tcatgcttgc ggacatcctg gcatacttga atgaaaaagg tacattcgag gagattgccg
     3361 aagtggcaga ctacaataaa aaagtcttca ttgcattagg acttgacgaa agccgcgcca
     3421 aattcgtctt gggtagcgaa tatgagctga gccgtgacta cgtgttagac gtgttaaaga
     3481 tggcccgtat cactacgctt aatcgtgcgc gtcgcagcat ggatgaggtc agccgccgca
     3541 aggaagatcc aatggtttcc caaatgattt acccgcttat gcaggcactg acgggggctc
     3601 atttaggagt ggaccttgcc gtaggtggaa tcgaccaacg taagatccat atgctggcgc
     3661 gcgaaaacct gcctcgcttg ggatatagta gcccggtgtg tttacatacg ccgattctgg
     3721 tggggttaga cggacaaaag atgagtagca gtaagggaaa ttacattagt gttcgtgatc
     3781 ctccagaaga ggtggagcgt aaaatccgta aagcttattg cccggcagga gtggtcgagg
     3841 agaatccgat tctggatatt gccaagtatc acattttgcc ccgttttggt aagatcgtag
     3901 tagagcgcga tgctaaggtg ggaggggacg ttgaatacgc gagtttcgag gaattggcag
     3961 aagactttaa gtctggtcag ctgcatccag ggaggttaaa gatcgcggtc gctaaatatt
     4021 tgaatatgct gttggaggat gcccgtaaac gtctgggcgt gtctgtttaa gtcttccact
     4081 gctagcatta acattagcga ccggcaagca ggatagtaag gaggtttttt ggaggacatc
     4141 ctgcagtttc aaacgctaaa ttgcctgatg cgctacgctt atcaggccta catgatctct
     4201 gcaatatatt gagtttgcgt gcttttgtag gccggataag gcgttcacgc cgcatccggc
     4261 aagaaacagc aaacaatcca aaacgccgcg ttcggcggtc gacacagatg taggtgttcc
     4321 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgcttg
     4381 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ctgccggcac
     4441 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
     4501 ccgcgcccac cggaaggagc taccggcagc ggtgcggact gttgtaactc agaataagaa
     4561 atgaggccgc tcatggcgtt ctgttgcccg tctcactggt gaaaagaaaa acaaccctgg
     4621 cgccgcttct ttgagcgaac gatcaaaaat aagtggcgcc ccatcaaaaa aatattctca
     4681 acataaaaaa ctttgtgtaa tacttgtaac gctagatctg ggggacggtc cggcgaccag
     4741 cgggtctcga aaacctagcc ttgcggggtt cgacaccccg gtctctcgcc aaattttgca
     4801 cccagcaaac ttggtacgta aacgcatcgt cccgccctag ctcagaggta gagcgtgctt
     4861 ctctaaaaag catggtcccc ggttcaaatc ctgggggcgg gaccaactag tatccttagc
     4921 gaaagctaag gatttttttt aagcttggca ctggccgtcg ttttacaacg tcgtgactgg
     4981 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagacgc
     5041 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     5101 accgccgccg caag
//
