Basic Vector Information
- Vector Name:
- pMB1_1R26PylRS.CbzK-AfTyrRS.p-I-Phe
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5114 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Robertson WE, Funke LFH., de la Torre D, Fredens J
pMB1_1R26PylRS.CbzK-AfTyrRS.p-I-Phe 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_1R26PylRS.CbzK-AfTyrRS.p-I-Phe vector Sequence
LOCUS 62056_16540 5114 bp DNA circular SYN 12-JUN-2021
DEFINITION Cloning vector pMB1_1R26PylRS.CbzK/AfTyrRS.p-I-Phe, complete
sequence.
ACCESSION MW879725
VERSION .
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 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..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=tyrS
/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"
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