pMB1_MmPylRS-1R26PylRS.CbzK vector (V015699)

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"