pMB1_1R26PylRS.CbzK-AfTyrRS.p-I-Phe vector (V015808)

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"