pBAD_sfGFP-3TCG4TCG_MmPylRS vector (V016067)

Basic Vector Information

Vector Name:
pBAD_sfGFP-3TCG4TCG_MmPylRS
Antibiotic Resistance:
Apramycin
Length:
6853 bp
Type:
Cloning vector
Replication origin:
p15A ori
Source/Author:
Robertson WE, Funke LFH., de la Torre D, Fredens J
Promoter:
araBAD

pBAD_sfGFP-3TCG4TCG_MmPylRS vector Map

pBAD_sfGFP-3TCG4TCG_MmPylRS6853 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600glnSPyrrolysine--tRNA ligaseFLAGlpp promotertRNA-SerrrnC terminator; rrnC termrrnB T1 terminatorp15A oritet promoterAmpR promoterApmRaraCaraBAD promotersuperfolder GFP6xHisMyc6xHis

pBAD_sfGFP-3TCG4TCG_MmPylRS vector Sequence

LOCUS       V016067                 6853 bp    DNA     circular SYN 12-JUN-2021
DEFINITION  Exported.
ACCESSION   V016067
VERSION     V016067
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6853)
  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 6853)
  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 6853)
  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..6853
                     /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             5992..6702
                     /label="superfolder GFP"
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     CDS             6709..6726
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             6752..6781
                     /label="Myc"
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             6797..6814
                     /label="6xHis"
                     /note="6xHis affinity tag"

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