Basic Vector Information
- Vector Name:
- pMB1_MmPylRS-1R26PylRS.CbzK
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5524 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Robertson WE, Funke LFH., de la Torre D, Fredens J
pMB1_MmPylRS-1R26PylRS.CbzK vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pMB1_MmPylRS-1R26PylRS.CbzK vector Sequence
LOCUS 62056_16545 5524 bp DNA circular SYN 12-JUN-2021
DEFINITION Cloning vector pMB1_MmPylRS/1R26PylRS.CbzK, complete sequence.
ACCESSION MW879726
VERSION .
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 SGRef: number: 1; type: "Journal Article"; journalName: "Science
(2021) In press"
COMMENT 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"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
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=pylS
/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"
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